Coiled-coil domain-containing protein 152 (CCDC152) has been identified as a gene involved in cell cycle regulation, cellular proliferation and migration. Overexpression of CCDC152 reduced cell proliferation and migration through the JAK2/STAT signaling pathway [
].
This entry represents mediator of RNA polymerase II transcription subunit 2 (Med2) from plants.The Mediator complex is a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. The Mediator complex, having a compact conformation in its free form, is recruited to promoters by direct interactions with regulatory proteins and serves for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. On recruitment the Mediator complex unfolds to an extended conformation and partially surrounds RNA polymerase II, specifically interacting with the unphosphorylated form of the C-terminal domain (CTD) of RNA polymerase II. The Mediator complex dissociates from the RNA polymerase II holoenzyme and stays at the promoter when transcriptional elongation begins [
].
Histones can be reversibly acetylated on several lysine residues.
Regulation of transcription is caused in part by thismechanism. Histone deacetylases catalyse the removal
of the acetyl group. Histone deacetylases, acetoin utilization proteins and acetylpolyamine amidohydrolases are all members of this ancient protein superfamily []. Histone deacetylases (HDA), acetoin utilisation proteins (ACUC) and
acetylpolyamine amidohydrolases (APHA) form an ancient protein superfamilyand are all believed to catalyse the deacetylation of a substrate [
]. HDAs are found in eukaryotes, from yeast to humans; ACUCs are found in eubacteria; and APHAs occur in archaeal bacteria, eubacteria and some eukaryotic organisms []. HDA nomenclature is confusing: the family can also be identified by the codes RPD, HOS, HD and HDAC, regardless ofspecific sequence similarities. HDAs may have evolved at least twice from APHAs, the more recent gene duplication event resembling APHAs more closely [
].Disrupting the expression of ACUC results in diminished growth of bacteria on acetoin and butanediol. Acetoin is a bacterial fermentation product that can be converted to acetate via the butanediol cycle in the absence of other carbon sources; its degradation is believed to be caused by deacetylation mediated by ACUC [
].
BROX has a Bro1 domain-like sequence and a C-terminal thioester-linkage site of isoprenoid lipid (CAAX motif). It interacts with CHMP4, a component of endosomal sorting complex required for transport III [
].
This entry represents the non classical export protein 1 family. Family members are Involved in a novel pathway of export of proteins that lack a cleavable signal sequence [
].
Geminiviruses are characterised by a genome of circular single-stranded DNA encapsidated in twinned (geminate) quasi-isometric particles, from which the group derives its name [
]. Most geminiviruses can be divided into two subgroups on the basis of host range and/or insect vector: i.e. those that infect dicotyledenous plants and are transmitted by the same whitefly species, and those that infect monocotyledenous plants and are transmitted by different leafhopper vectors. The genomes of the whitefly-transmitted African cassava mosaic virus, Tomato golden mosaic virus (TGMV) and Bean golden mosaic virus (BGMV) possess a bipartite genome. By contrast, only a single DNA component has been identified for the leafhopper-transmitted Maize streak virus (MSV) and Wheat dwarf virus (WDV) [,
]. Beet curly top virus (BCTV), and Tobacco yellow dwarf virus belong to a third possible subgroup. Like MSV and WDV, BCTV is transmitted by a specific leafhopper species, yet like the whitefly-transmitted geminiviruses it has a host range confined to dicotyledenous plants.Sequence comparison of the whitefly-transmitted Squash leaf curl virus (SqLCV) and Tomato yellow leaf curl virus (TYLCV) with the genomic components of TGMV and BGMV reveals a close evolutionary relationship [
,
,
]. Amino acid sequence alignments of Potato yellow mosaic virus (PYMV) proteins with those encoded by other geminiviruses show that PYMV is closely related to geminiviruses isolated from the New World, especially in the putative coat protein gene regions []. Comparison of MSV DNA-encoded proteins with those of other geminiviruses infecting monocotyledonous plants, including Panicum streak virus [] and Miscanthus streak virus (MiSV) [], reveal high levels of similarity.
Geminiviruses are characterised by a genome of circular single-stranded DNA encapsidated in twinned (geminate) quasi-isometric particles, from which the group derives its name [
]. Most geminiviruses can be divided into two subgroups on the basis of host range and/or insect vector: i.e. those that infect dicotyledenous plants and are transmitted by the same whitefly species, and those that infect monocotyledenous plants and are transmitted by different leafhopper vectors. The genomes of the whitefly-transmitted African cassava mosaic virus, Tomato golden mosaic virus (TGMV) and Bean golden mosaic virus (BGMV) possess a bipartite genome. By contrast, only a single DNA component has been identified for the leafhopper-transmitted Maize streak virus (MSV) and Wheat dwarf virus (WDV) [,
]. Beet curly top virus (BCTV), and Tobacco yellow dwarf virus belong to a third possible subgroup. Like MSV and WDV, BCTV is transmitted by a specific leafhopper species, yet like the whitefly-transmitted geminiviruses it has a host range confined to dicotyledenous plants.Sequence comparison of the whitefly-transmitted Squash leaf curl virus (SqLCV) and Tomato yellow leaf curl virus (TYLCV) with the genomic components of TGMV and BGMV reveals a close evolutionary relationship [
,
,
]. Amino acid sequence alignments of Potato yellow mosaic virus (PYMV) proteins with those encoded by other geminiviruses show that PYMV is closely related to geminiviruses isolated from the New World, especially in the putative coat protein gene regions []. Comparison of MSV DNA-encoded proteins with those of other geminiviruses infecting monocotyledonous plants, including Panicum streak virus [] and Miscanthus streak virus (MiSV) [], reveal high levels of similarity.
A PB1 domain is present in NIN like proteins (NLP), key transcription factors in a process of establishment of symbiosis between legumes and nitrogen fixing bacteria (Rhizobium) [
] and nitrate signalling in other plants []. NLPs carry a PB1 domain at their C terminus [].
Activity-dependent neuroprotector homeobox protein
Type:
Family
Description:
This entry includes a group of homeobox proteins, including ADNP and ADNP2 [
]. ADNP regulates the expression of >400 genes during embryonic development, vital for mammalian brain formation [
,
]. It has been linked to neurodegenerative diseases and has been shown to play an important roles in autophagy. ADNP has also been associated with the pathophysiology of schizophrenia []. ADNP de novo mutations in humans result in a syndromic form of autism-like spectrum disorder (ASD), including cognitive and motor deficits, the ADNP syndrome (Helsmoortel-Van Der Aa) [].
This entry represents the main structural domain of She2.She2 is a RNA binding protein which binds to RNA via a helical hairpin. The protein is required for the actin dependent transport of ASH1 mRNA in yeast, a form of mRNP translocation [
]. She2 contains a globular domain consisting of a bundle of five α-helices [].
Many flagellar proteins are exported by a flagellum-specific export pathway. Attempts have been made to characterise
the apparatus responsible for this process, by designing assays to screen for mutants with export defects.Experiments involving filament removal from temperature-sensitive flagellar mutants of Salmonella typhimurium have
shown that, while most mutants were able to regrow filaments, flhA, fliH, fliI and fliN mutants showed no or greatlyreduced regrowth. This suggests that the corresponding gene products are involved in the process of flagellum-specific export [
]. The sequence of fliH has been deduced and shown to encode a protein of molecular massof 25,782 Da.
This is a domain found at the C-terminal of Wadjet protein JetD from Bacillus cereus. JetD is a component of the antiplasmid transformation system Wadjet type I, composed of JetA, JetB, JetC and JetD. Expression of Wadjet type I in B.subtilis (strain BEST7003) reduces the transformation efficiency of plasmid pHCMC05 [
]. This domain is structurally similar to DNA topoisomerases, therefore, it may be a DNA-binding domain.
This superfamily consists of bacterial (and chloroplast) examples of the ribosomal small subunit protein S20. Bacterial ribosomal protein S20 forms part of the 30S ribosomal subunit, and interacts with 16S rRNA.Structurally, the ribosomal protein S20 has a spectrin repeat-like fold, which consists of three helices, a close bundle fold and left-handed twist going up-and-down.
This entry covers a section of the β-propeller found at the N-terminal of NOL11 [
].NOL11 is a nucleolar protein and a component of the human ribosomal small subunit (SSU) processome. It is required for the early stages of ribosome biogenesis in humans [
]. It interacts with the C-terminal region of the known t-UTP/UTPA subcomplex member, hUTP4/Cirhin, the protein mutated in North American Indian childhood cirrhosis (NAIC) []. In Xenopus, it is required for optimal rDNA transcription and craniofacial development [].
Rix1 is a nucleoplasmic particle involved in rRNA processing/ribosome assembly [
,
]. It associates with two other proteins, Ipi1 and Ipi3, to form the RIX1 complex that allows Rea1 - the AAA ATPase - to associate with the 60S ribosomal subunit. More than 170 assembly factors are involved in the construction and maturation of yeast ribosomes, and after these factors have completed their function they need to be released from the pre-ribosomes. Rea1 induces the release of the assembly protein complex in a mechanical fashion []. This family is usually associated with NUC202, .
The cloacin immunity protein complexes with cloacin in equimolar quantities and inhibits it by binding with high affinity to the cloacin C-terminal catalytic domain. The immunity protein is relatively small, containing 85 amino acids.An extra ribosome binding site has been found to precede the immunity gene on the polycistronic Clo DF13 mRNA [
], which perhaps accounts for the fact that, in cloacinogenic cells, more immunity protein than cloacin is synthesised []. Comparison of the complete amino acid sequence of the Clo DF13 immunity protein with that of the Col E3 and Col E6 immunity proteins reveals extensive similarities in primary structure, although Col E3 and Clo DF13 immunity proteins are exchangeable only to a low extent in vivoand
in vitro[
].The cloacin structure has a beta(2)-α-β(2) fold with antiparallel β-sheet.
Autophagy is generally known as a process involved in the degradation of bulk cytoplasmic components that are sequestered into double-membrane vesicles to form the autophagosome. The contents of the autophagosome are delivered to the degradative organelle, the lysosome/vacuole, for breakdown and eventual recycling of the resulting macromolecules. In contrast to autophagy, the cytoplasm to vacuole transport (Cvt) pathway is a highly selective process that involves the sequestration of at least two specific cargos that are resident vacuolar hydrolases: aminopeptidase I (Ape1) and alpha-mannosidase (Ams1). These proteins are sequestered within a double-membrane vesicle, termed a Cvt vesicle. The Cvt vesicle is fairly consistent in size, and is much smaller than the autophagosome, being 140-160 nm in diameter.The genes involved in autophagy in yeast are termed autophagy-related (Atg) genes. Autophagy and the Cvt pathway are topologically and mechanistically similar and share most of the Atg components []. The precursor of Ape1 (prApe1) is sequestered within either Cvt vesicles or autophagosomes, depending on the nutrient conditions, and delivered to the vacuole.Atg19 is a peripheral membrane protein with differing binding sites for both Ape1 and Ams1. Atg19 plays a central role in cargo sorting and transport to the vacuole by linking Ams1 and prApe1 to Atg8 and Atg11 [
].This entry represents the C-terminal region of Atg19 and homologue Atg34 (Yol083wp) [
,
]. The Atg8-binding region is thought to be in the very C-terminal residues [].
The fliD operon of several bacteria consists of three flagellar genes, fliD, fliS, and fliT, and is transcribed in this order [
]. In Bacillus subtilis the operon encoding the flagellar proteins FliD, FliS, and FliT is sigma D-dependent [].FliS is a flagellin-specific T3S chaperone that binds in 1:1 stoichiometry to the C-terminal region of flagellin and facilitates its export, preventing polymerization in the cytoplasm [
,
,
]. The fliS gene is necessary for flagellation. In Aeromonas, deletion of this gene (called flaJ) resulted in the complete loss of motility, flagellin expression, and adherence [].Structurally, FliS can form closed, open and helix-swapped bundles.
Flaviviruses encode a single polyprotein. This is cleaved into
three structural and seven non-structural proteins. The NS4Aprotein is small and poorly conserved among the Flaviviruses.
NS4A contains multiple hydrophobic potential membrane spanningregions [
]. NS4A has only been found in cells infected by Kunjin virus [].
Daunorubicin resistance protein C confers the function of daunorubicin resistance. The protein seems to share strong sequence similarity to UvrA proteins, which are involved in excision repair of DNA. Disruption of drrC gene showed increased sensitivity upon exposure to duanorubicin, however it failed to complement uvrA mutants to exposure to UV irradiation. The mechanism on how it confers duanomycin resistance is unclear, but has been suggested to be different from DrrA and DrrB which are antiporters.
This entry represents a cell division protein, designated SepF, which is conserved in Gram-positive bacteria. SepF accumulates at the cell division site in an FtsZ-dependent manner and is required for proper septum formation [
,
]. This protein uses an amphipathic helix for membrane binding and polymerises into large rings that bundle FtsZ filaments []. Mutants are viable but the formation of the septum is much slower and occurs with a very abnormal morphology. This entry also includes archaeal related proteins, such as SepF from Haloferax volcanii [].
This family of bacterial proteins is functionally uncharacterised. Proteins in this family are typically between 116 and 146 amino acids in length. PrgI is found encoded on plasmids of Enterococcus faecalis, its function is not known.
This entry represents a group of WYL domain containing proteins. WYL is named for three conserved amino acids found in a subset of domains of this superfamily. These residues are not strongly conserved throughout the family. Rather, the conservation pattern includes four basic residues and a position often occupied by a cysteine [
], which are predicted to line a ligand-binding groove typical of the Sm-like SH3 β-barrels. It is predicted to be a ligand-sensing domain that could bind negatively charged ligands, such as nucleotides or nucleic acid fragments, to regulate CRISPR-Cas and other defense systems such as the abortive infection AbiG system [].
Urease and other nickel metalloenzymes are synthesised as precursors devoid of the metalloenzyme active site. These precursors then undergo a complex post-translational maturation process that requires a number of accessory proteins.Members of this group are nickel-binding proteins required for urease metallocentre assembly [
]. They are believed to function as metallochaperones to deliver nickel to urease apoprotein [,
]. It has been shown by yeast two-hybrid analysis that UreE forms a dimeric complex with UreG in Helicobacter pylori 26695 []. The UreDFG-apoenzyme complex has also been shown to exist [
,
] and is believed to be, with the addition of UreE, the assembly system for active urease []. The complexes, rather than the individual proteins, presumably bind to UreB via UreE/H recognition sites.The structure of Klebsiella aerogenes UreE reveals a unique two-domain architecture with one domain structurally related to a heat shock protein and the second to the Atx1 copper metallochaperone [
,
]. Significantly, the metal-binding sites in UreE and Atx1 are distinct in location and types of residues despite the relationship between these proteins and the mechanism for UreE activation of urease is proposed to be different from the thiol ligand exchange mechanism used by the copper metallochaperones.Most members of this group contain a histidine-rich C-terminal motif that is involved in, but not solely responsible for, binding nickel ions in K. aerogenes UreE [
]. However, internal ligands, not the histidine residues at the C terminus, are necessary for UreE to assist in urease activation in K. aerogenes [], even though the truncated protein lacking the His-rich region binds two nickel ions instead of six. In H. pylori and some other organisms, the terminal histidine-rich binding sites are absent, but the internal histidine sites are present, and the latter probably function as nickel donors.
Proteins in this family are copper ion-binding proteins, including copper chaperone CopZ and COP-associated protein (COPP). They contain a conserved 30-residue HMA domain, which has been found in a number of other heavy metal transport or detoxification proteins [
]. CopZ is a chaperone that serves for the intracellular sequestration and transport of Cu+, and delivers copper ions to the copper-transporting ATPase CopA []. COPP is part of a cation-transporting system which is associated with copper export out of the cells [].
PriB is a component of the preprimosomal complex. The preprimosomal complex is composed of one monomer of PriC and DnaT, two monomers of PriA, two dimers of PriB and one hexamer of DnaB. Upon transient interaction with DnaG it forms the primosome [
].PriB binds single-stranded DNA at the primosome assembly site (PAS) [
,
,
]. Assembly of the primosome is initiated by interactions of PriA and PriB with ssDNA and the PAS [].
Ribosomes are the particles that catalyse mRNA-directed protein synthesis in all organisms. The codons of the mRNA are exposed on the ribosome to allow tRNA binding. This leads to the incorporation of amino acids into the growing polypeptide chain in accordance with the genetic information. Incoming amino acid monomers enter the ribosomal A site in the form of aminoacyl-tRNAs complexed with elongation factor Tu (EF-Tu) and GTP. The growing polypeptide chain, situated in the P site as peptidyl-tRNA, is then transferred to aminoacyl-tRNA and the new peptidyl-tRNA, extended by one residue, is translocated to the P site with the aid the elongation factor G (EF-G) and GTP as the deacylated tRNA is released from the ribosome through one or more exit sites [
,
]. About 2/3 of the mass of the ribosome consists of RNA and 1/3 of protein. The proteins are named in accordance with the subunit of the ribosome which they belong to - the small (S1 to S31) and the large (L1 to L44). Usually they decorate the rRNA cores of the subunits. Many ribosomal proteins, particularly those of the large subunit, are composed of a globular, surfaced-exposed domain with long finger-like projections that extend into the rRNA core to stabilise its structure. Most of the proteins interact with multiple RNA elements, often from different domains. In the large subunit, about 1/3 of the 23S rRNA nucleotides are at least in van der Waal's contact with protein, and L22 interacts with all six domains of the 23S rRNA. Proteins S4 and S7, which initiate assembly of the 16S rRNA, are located at junctions of five and four RNA helices, respectively. In this way proteins serve to organise and stabilise the rRNA tertiary structure. While the crucial activities of decoding and peptide transfer are RNA based, proteins play an active role in functions that may have evolved to streamline the process of protein synthesis. In addition to their function in the ribosome, many ribosomal proteins have some function 'outside' the ribosome [
,
].A number of eukaryotic and archaebacterial ribosomal proteins can be grouped
on the basis of sequence similarities []. One of these families consists of:Mammalian L15.Insect L15.Plant L15.Yeast YL10 (L13) (Rp15r).Archaebacterial L15e.These proteins have about 200 amino acid residues.This entry represents the archaeal L15e proteins.
Pathogenic members of the flavivirus family [E1], including West Nile Virus(WNV) and Dengue Virus (DV), are growing global threats for which there are no
specific treatments. The genome encodes three structural proteins found inthe mature virion (C, prM, and E) and seven "nonstructural"(i.e., not part of
the virion architecture) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5).Full-length NS3 is a bifunctional protein. The N-terminal 175 residues
comprise a chymotrypsin-like protease, "NS3pro", while the C-terminal portion
is a helicase ("NS3hel"). The NS2B protein, which is located in the
polypeptide precursor immediately upstream of the NS3pro domain, functions asthe cofactor for NS3pro. A 35-48 residue central portion is required for
protease activity in vitro, while N- and C-terminal flanking hydrophobicregions are predicted to anchor the NS2B-NS3 complex into the host endoplasmic
reticulum membrane. The two component flaviviral enzyme NS2B-NS3 cleaves theviral polyprotein precursor within the host cell, a process that is required
for viral replication [,
,
]. The NS3pro domain forms peptidase family S7(flavivirin family) of clan PA [E2].The NS3pro has a classical serine protease catalytic triad (His, Asp, and
Ser). The enzymatic activity of NS3pro is enhanced by interacting with thecentral 40 amino acid of NS2B which acts as an essential cofactor. The NS3pro
domain has an overall structure of two barrels made of six beta sheets each,with the active site located in the cleft between the barrels. The NS2B
hydrophilic core cofactor contributes one of the N-terminal beta sheets [,
,
].This entry represents a domain cover the entire flavivirus NS2B and NS3pro domains.
This entry represents a group of fungal proteins, including Pup1 from Candida glabrata. Pup1 is a mitochondrial protein that contributes to the enhanced virulence of C.glabrata strains that acquired azole resistance [
].
There is currently no experimental data for members of this group or their homologues, nor do they exhibit features indicative of any function. Members of this entry are mainly found in proteobacteria.
A number of eukaryotic and archaebacterial ribosomal proteins can be grouped in this superfamily of ribosomal proteins, S17e (RPS17e). They include, vertebrate, Drosophila and Neurospora crassa (crp-3) S17's as well as yeast S17a (RP51A) and S17b (RP51B) and archaebacterial S17e [
,
,
].RPS17e has a DNA/RNA-binding 3-helical bundle fold, which consists of three helices with a close or partly open bundle and right-handed twist going up-and down.
GerPE is required for the formation of functionally normal spores. It could be involved in the establishment of a normal spore coat structure and/or permeability, which allows the access of germinants to their receptor [
].
Members of this superfamily are essential for gametocytogenesis in Plasmodium falciparum. They contain a fold composed of two pseudo dyad-related repeats of the helix-turn-helix motif, serving as a platform for RNA and Src homology-3 (SH3) binding [
].
This entry consists of bacterial uncharacterised proteins. The structure of one of the proteins has been solved and it adopts a beta barrel-like structure.
Glycolipid transfer protein (GLTP) is a cytosolic protein that catalyses the intermembrane transfer of glycolipids such as glycosphingolipids, glyceroglycolipids, and possibly glucosylceramides, but not of phospholipids. The GLTP protein consists of a single domain with a multi-helical structure consisting of two layers of orthogonally packed helices [
,
]. The GLTP domain is also found in trans-Golgi network proteins involved in Golgi-to-cell-surface membrane traffic [
].
Ribosomes are the particles that catalyse mRNA-directed protein synthesis in all organisms. The codons of the mRNA are exposed on the ribosome to allow tRNA binding. This leads to the incorporation of amino acids into the growing polypeptide chain in accordance with the genetic information. Incoming amino acid monomers enter the ribosomal A site in the form of aminoacyl-tRNAs complexed with elongation factor Tu (EF-Tu) and GTP. The growing polypeptide chain, situated in the P site as peptidyl-tRNA, is then transferred to aminoacyl-tRNA and the new peptidyl-tRNA, extended by one residue, is translocated to the P site with the aid the elongation factor G (EF-G) and GTP as the deacylated tRNA is released from the ribosome through one or more exit sites [
,
]. About 2/3 of the mass of the ribosome consists of RNA and 1/3 of protein. The proteins are named in accordance with the subunit of the ribosome which they belong to - the small (S1 to S31) and the large (L1 to L44). Usually they decorate the rRNA cores of the subunits. Many ribosomal proteins, particularly those of the large subunit, are composed of a globular, surfaced-exposed domain with long finger-like projections that extend into the rRNA core to stabilise its structure. Most of the proteins interact with multiple RNA elements, often from different domains. In the large subunit, about 1/3 of the 23S rRNA nucleotides are at least in van der Waal's contact with protein, and L22 interacts with all six domains of the 23S rRNA. Proteins S4 and S7, which initiate assembly of the 16S rRNA, are located at junctions of five and four RNA helices, respectively. In this way proteins serve to organise and stabilise the rRNA tertiary structure. While the crucial activities of decoding and peptide transfer are RNA based, proteins play an active role in functions that may have evolved to streamline the process of protein synthesis. In addition to their function in the ribosome, many ribosomal proteins have some function 'outside' the ribosome [
,
].Members of this protein family are the archaeal ribosomal protein L6P.
This entry represents the kinase-like domain found in the Rhoptry (ROP) pseudokinases from the intracellular parasite Toxoplasma gondii [
]. ROP2, ROP4 and ROP8 have a typical bi-lobed protein kinase fold, but lack catalytic activity [].ROP4 is localised to the rhoptries, secretory organelles at the apical end of the parasite, and is secreted from the parasite during host cell invasion [
]. ROP2 mediates an association between the parasitophorous vacuole (PV) and host cell mitochondria by inserting into the parasitophorous vacuole membrane (PVM) []. It plays important roles in rhoptry biogenesis, parasite invasion, and intracellular replication []. During host invasion, ROP2 is released from the vacuole membrane and becomes phosphorylated in the infected cells [].
PINK1 is a mitochondrial serine/threonine kinase that acts as a sensor of mitochondrial damage [
]. It phosphorylates ubiquitin (Ub) and the Ub-like domain (UBL) of the E3 Ub ligase parkin at Ser65. The phosphorylation of Ub and the parkin UBL facilitates the transition from the autoinhibition state to the catalytically active state in parkin []. Upon mitochondrial damage, PINK1 accumulates on the outer mitochondrial membrane (OMM), where it may recruit and activate Parkin []. PINK1 has also beenshown to phosphorylate Miro, a component of the primary motor/adaptor complex that anchors kinesin to the mitochondrial surface. The phosphorylation of Miro activates proteasomal degradation of Miro in a Parkin-dependent manner [
]. PINK1 contains an N-terminal mitochondrial targeting sequence (MTS), a transmembrane domain (TM), a highly conserved serine/threonine kinase domain, and a C-terminal auto-regulatory domain. Mutations of the PINK1 and Parkin genes have been linked to familial Parkinson's disease (PD) [
].
TcpC is required for efficient conjugative transfer, localizing to the cell membrane independently of other conjugation proteins, where membrane localization is important for its function, oligomerization and interaction with the conjugation proteins TcpA, TcpH, and TcpG [
]. Its C-terminal domain is critical for interactions with these other conjugation proteins []. TcpC has low level sequence identity to proteins encoded by the conjugative transposon Tn916, which is responsible for a large proportion of the tetracycline resistance in different pathogens [].
This entry describes a subfamily of the B12 binding domain proteins that include corrinoid proteins specific to four different, mutually non-homologous enzymes of the genus Methanosarcina. Three of the four cognate enzymes (trimethylamine, dimethylamine, and monomethylamine methyltransferases) all have the unusual, ribosomally incorporated amino acid pyrrolysine at the active site. All act in systems in which a methyl group is transferred to the corrinoid protein to create methylcobalamin, from which the methyl group is later transferred elsewhere.
Members of the HesA/MoeB/ThiF family of proteins (
) include a number of members encoded in the midst of thiamine biosynthetic operons. This mix of known and putative ThiF proteins shows a deep split in phylogenetic trees. The Escherichia coli ThiF and MoeB proteins are seemingly more closely related than the E. coli ThiF and Campylobacter (for example) ThiF. This entry represents the divergent clade of putative ThiF proteins as found in Campylobacter.Members of this family are members of the superfamily of activating enzymes (E1) of the ubiquitin-like proteins [
]. The exact function of this family is unknown.
NS2A is a hydrophobic protein about 25kDa in size, which is cleaved from NS1 by a membrane
bound host protease []. NS2A has been found to associate with the dsRNA within the vesicle packages. It has also been found that NS2A associates with the known replicase
components and so NS2A has been postulated to be part of this replicase complex [].
Ribosomes are the particles that catalyse mRNA-directed protein synthesis in all organisms. The codons of the mRNA are exposed on the ribosome to allow tRNA binding. This leads to the incorporation of amino acids into the growing polypeptide chain in accordance with the genetic information. Incoming amino acid monomers enter the ribosomal A site in the form of aminoacyl-tRNAs complexed with elongation factor Tu (EF-Tu) and GTP. The growing polypeptide chain, situated in the P site as peptidyl-tRNA, is then transferred to aminoacyl-tRNA and the new peptidyl-tRNA, extended by one residue, is translocated to the P site with the aid the elongation factor G (EF-G) and GTP as the deacylated tRNA is released from the ribosome through one or more exit sites [,
]. About 2/3 of the mass of the ribosome consists of RNA and 1/3 of protein. The proteins are named in accordance with the subunit of the ribosome which they belong to - the small (S1 to S31) and the large (L1 to L44). Usually they decorate the rRNA cores of the subunits. Many ribosomal proteins, particularly those of the large subunit, are composed of a globular, surfaced-exposed domain with long finger-like projections that extend into the rRNA core to stabilise its structure. Most of the proteins interact with multiple RNA elements, often from different domains. In the large subunit, about 1/3 of the 23S rRNA nucleotides are at least in van der Waal's contact with protein, and L22 interacts with all six domains of the 23S rRNA. Proteins S4 and S7, which initiate assembly of the 16S rRNA, are located at junctions of five and four RNA helices, respectively. In this way proteins serve to organise and stabilise the rRNA tertiary structure. While the crucial activities of decoding and peptide transfer are RNA based, proteins play an active role in functions that may have evolved to streamline the process of protein synthesis. In addition to their function in the ribosome, many ribosomal proteins have some function 'outside' the ribosome [
,
].Ribosomal protein S9 is one of the proteins from the small ribosomal subunit. It belongs to the S9P family of ribosomal proteins which, on the basis of sequence similarities [
], groups bacterial; algal chloroplast; cyanelle and archaeal S9 proteins; and mammalian, plant, and yeast mitochondrial ribosomal S9 proteins. These proteins adopt a β-α-β fold similar to that found in numerous RNA/DNA-binding proteins, as well as in kinases from the GHMP kinase family [].This entry represents archaeal ribosomal S9 proteins.
This entry includes the outer capsid protein Mu1 and VP4. Mu1 is an outer capsid protein that acts as a reoviral penetration agent. Non-enveloped animal reoviruses must enter host cells by membrane penetration that does not involve membrane fusion, as they lack a viral membrane. Reoviruses are activated by proteolytic cleavage in the intestinal lumen, leading to infectious subviral particles. The core of the virus is coated by a layer of mu1 and sigma3 proteins. Proteases strip off sigma3 exposing mu1, which provides the membrane penetration machinery that perforates the membrane. In addition, N-terminal myristoylation of polypeptide Mu1 are required for site-specific cleavage to Mu1C in transfected cells [
]. Mu1 forms a trimer, where the three mu1 molecules are coiled around one another with a right-handed twist. The mu1 chain folds into four distinct domains: three intertwined, predominantly alpha helical domains and a jelly-roll β-sandwich []. VP4 nteracts with VP7 to form the outer icosahedral capsid with an incomplete T=13 symmetry, about 80 nm in diameter, and consisting of 200 VP4-VP7 trimers. Its myristoylated N-terminal peptide may be released in the endosome and involved in permeabilization and delivery of transcriptionally active viral particles into the host cell cytoplasm [
,
].
The NSS proteins are encoded in the S RNA from ssRNA negative-strand viruses [
]. The S RNA also codes for the nucleoprotein N. The two main products are read from overlapping reading frames in the viral complementary sequence.
Steroidogenic acute regulatory protein (StAR) facilitates the biosynthesis of steroid hormones by regulating the transfer of cholesterol from the outer to the inner mitochondrial membrane [
]. Mitochondrial fusion and ERK phosphorylation can direct StAR to the outer mitochondrial membrane to achieve a large number of steroid molecules per unit of StAR [,
].
This domain is found in eukaryotes, and is typically between 69 and 88 amino acids in length. It is found in association with
. It contains two conserved sequence motifs: YDW and PVR. It is found in Sox8 [
], Sox9 [] and Sox10 [] proteins, which have structural similarity. Sox proteins are involved in developmental processes.
This domain is found in bacteria, and is approximately 100 amino acids in length. It is found in association with a von Willebrand factor type A domain (
). It contains two conserved sequence motifs: STF and DVD and two completely conserved residues (E and N) that may be functionally important. It is found in uncharacterised protein YfbK.
Detailed sequence analysis shows that members of this family are distantly related to thermophilic adenylate cyclases (class 2) (
) and mammalian thiamine triphosphatases, and belong to the CYTH domain superfamily [
]. It has been predicted that CYTH domain proteins may play a central role in the interface between nucleotide and polyphosphate metabolism []. Based on the conservation of catalytic residues, it has been predicted that CYTH domains are likely to chelate two divalent cations, and exhibit a reaction mechanism that is dependent on two metal ions, analogous to nucleotide cyclases, polymerases and certain phosphoesterases []. It has also been suggested that the experimentally characterised members of the CYTH domain superfamily, namely adenylyl cyclase and thiamine triphosphatase, are secondary derivatives of proteins that performed an ancient role in polyphosphate and nucleotide metabolism [].
This entry represents a group of CRISPR-associated proteins belonging to the type III RAMP (Repair Associated Mysterious Proteins) family [
]. Members including in this family are Csm3, Csm5, Cmr6, Cmr4 and Cmr1. The CRISPR-Cas system is a prokaryotic defense mechanism against foreign genetic elements. The key elements of this defense system are the Cas proteins and the CRISPR RNA. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) are a family of DNA direct repeats separated by regularly sized non-repetitive spacer sequences that are found in most bacterial and archaeal genomes [
]. CRISPRs appear to provide acquired resistance against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain sequences complementary to antecedent mobile elements and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA).The defense reaction is divided into three stages. In the adaptation stage, the invader DNA is cleaved, and a piece of it is selected to be integrated as a new spacer into the CRISPR locus, where it is stored as an identity tag for future attacks by this invader. During the second stage (the expression stage), the CRISPR RNA (pre-crRNA) is transcribed and subsequently processed into the mature crRNAs. In the third stage (the interference stage), Cas proteins, together with crRNAs, identify and degrade the invader [
,
,
].The CRISPR-Cas systems have been sorted into three major classes. In CRISPR-Cas types I and III, the mature crRNA is generally generated by a member of the Cas6 protein family. Whereas in system III the Cas6 protein acts alone, in some class I systems it is part of a complex of Cas proteins known as Cascade (CRISPR-associated complex for antiviral defense). The Cas6 protein is an endoribonuclease necessary for crRNA production whereas the additional Cas proteins that form the Cascade complex are needed for crRNA stability [
].
This entry represents a family of highly hydrophobic, uncharacterised predicted integral membrane proteins found almost entirely in low-GC Gram-positive bacteria, although a member is also found in Aquifex aeolicus.
This domain is found in the C terminus of the paired-box protein 2, which is a transcription factor involved in embryonic development and organogenesis. It is approximately 110 amino acids in length and is found in association with
.
The pestivirus NS2 peptidase is responsible for single cleavage between NS2 and NS3 of the Bovine viral diarrhea virus 1 polyprotein, a cleavage that is correlated with cytopathogenicity [
]. The peptidase is activated by its interaction with 'J-domain protein interacting with viral protein' Jiv [].
The CRISPR-Cas system is a prokaryotic defense mechanism against foreign genetic elements. The key elements of this defense system are the Cas proteins and the CRISPR RNA. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) are a family of DNA direct repeats separated by regularly sized non-repetitive spacer sequences that are found in most bacterial and archaeal genomes [
]. CRISPRs appear to provide acquired resistance against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain sequences complementary to antecedent mobile elements and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA).The defense reaction is divided into three stages. In the adaptation stage, the invader DNA is cleaved, and a piece of it is selected to be integrated as a new spacer into the CRISPR locus, where it is stored as an identity tag for future attacks by this invader. During the second stage (the expression stage), the CRISPR RNA (pre-crRNA) is transcribed and subsequently processed into the mature crRNAs. In the third stage (the interference stage), Cas proteins, together with crRNAs, identify and degrade the invader [
,
,
].The CRISPR-Cas systems have been sorted into three major classes. In CRISPR-Cas types I and III, the mature crRNA is generally generated by a member of the Cas6 protein family. Whereas in system III the Cas6 protein acts alone, in some class I systems it is part of a complex of Cas proteins known as Cascade (CRISPR-associated complex for antiviral defense). The Cas6 protein is an endoribonuclease necessary for crRNA production whereas the additional Cas proteins that form the Cascade complex are needed for crRNA stability [
]. This entry represents a minor branch of the Cas2 family of CRISPR-associated proteins. Cas2 is one of four protein families (Cas1 to Cas4) that are associated with CRISPR elements and always occur near a repeat cluster, usually in the order cas3-cas4-cas1-cas2. The function of Cas2 (and Cas1) is unknown. Cas3 proteins appear to be helicases while Cas4 proteins resemble RecB-type exonucleases, suggesting that these genes are involved in DNA metabolism or gene expression [
].
This entry includes Vip3A and related proteins, such as Vip3B and Vip3C. They are insecticidal proteins produced during the vegetative growth phase of B. thuringiensis. Vip3A contains a conserved N-terminal domain and a highly variable C-terminal region. After ingestion of the toxin by the larva, Vip3A is activated through proteolytic process by trypsin-like proteases [
].
This domain family is typically between 101 and 138 amino acids in length and is found in subunit Crm2 of the CRISPR RNA-Cas protein complex (Cmr complex) [
].
Geminiviruses are characterised by a genome of circular single-stranded DNA encapsidated in twinned (geminate) quasi-isometric particles, from which the group derives its name [
]. Most geminiviruses can be divided into two subgroups on the basis of host range and/or insect vector: i.e. those that infect dicotyledenous plants and are transmitted by the same whitefly species, and those that infect monocotyledenous plants and are transmitted by different leafhopper vectors. The genomes of the whitefly-transmitted African cassava mosaic virus, Tomato golden mosaic virus (TGMV) and Bean golden mosaic virus (BGMV) possess a bipartite genome. By contrast, only a single DNA component has been identified for the leafhopper-transmitted Maize streak virus (MSV) and Wheat dwarf virus (WDV) [,
]. Beet curly top virus (BCTV), and Tobacco yellow dwarf virus belong to a third possible subgroup. Like MSV and WDV, BCTV is transmitted by a specific leafhopper species, yet like the whitefly-transmitted geminiviruses it has a host range confined to dicotyledenous plants.Sequence comparison of the whitefly-transmitted Squash leaf curl virus (SqLCV) and Tomato yellow leaf curl virus (TYLCV) with the genomic components of TGMV and BGMV reveals a close evolutionary relationship [
,
,
]. Amino acid sequence alignments of Potato yellow mosaic virus (PYMV) proteins with those encoded by other geminiviruses show that PYMV is closely related to geminiviruses isolated from the New World, especially in the putative coat protein gene regions []. Comparison of MSV DNA-encoded proteins with those of other geminiviruses infecting monocotyledonous plants, including Panicum streak virus [
] and Miscanthus streak virus (MiSV) [], reveal high levels of similarity.Geminiviruses contain three ORFs (designated AL1, AL2, and AL3) that
overlap and are specified by multiple polycistronic mRNAs. The AL3 protein comprises approximately 0.05% of the cellular proteins and is present in the soluble and organelle fractions [
]. AL3 may form oligomers [
]. Immunoprecipitation of AL3 in a baculovirus expression system extracts expressing both AL1 and AL3 showed
that the two proteins also complex with each other. The AL3 protein is involved in viral replication.
Gas vesicles are small, hollow, gas filled protein structures found in several cyanobacterial and archaebacterial
microorganisms []. They allow the positioning of the bacteria at the favourable depth for growth.Gas vesicles are hollow cylindrical tubes, closed by a hollow, conical cap at each end. Both the conical end
caps and central cylinder are made up of 4-5 nm wide ribs that run at right angles to the long axis of thestructure. Gas vesicles seem to be constituted of two different protein components, GVPa and GVPc. GVPa, a
small protein of about 70 amino acid residues, is the main constituent of gas vesicles and form the essentialcore of the structure. The sequence of GVPa is extremely well conserved. GvpJ and GvpM, two proteins encoded
in the cluster of genes required for gas vesicle synthesis in the archaebacteria Halobacterium salinarium andHalobacterium mediterranei (Haloferax mediterranei), have been found [
] to be evolutionary related to GVPa. The exact functionof these two proteins is not known, although they could be important for determining the shape determination
gas vesicles. The N-terminal domain of Aphanizomenon flos-aquae protein GvpA/J is also related to GVPa.
This family consists of uncharacterised proteins in:
Caulobacter crescentus CB15, Bdellovibrio bacteriovorus HD100, Synechococcus sp. (strain WH8102), Silicibacter pomeroyi DSS-3, and Hyphomonas neptunium (strain ATCC 15444). The context of nearby genes differs substantially between members and does not point to any specific biological role.
This entry represents Ycf66, a protein that is restricted to the chloroplasts of simple plants and algae. It is also found in the cyanobacteria. The function is unknown. As the family is exclusively found in phototrophic organisms it may play a role in photosynthesis.
This family of proteins includes Gp5 from bacteriophage P22 and related proteins. Gp5 is involved in the formation of the pro-capsid shells in the bacteriophage. In total, there are 415 molecules of the coat protein which are arranged in an icosahedral shell [
].
This family consists of small hypothetical proteins, about 100 amino acids in length. The family includes five members (three in tandem) in Pseudomonas aeruginosa PAO1, and also in Pseudomonas putida (strain KT2440), four in Pseudomonas syringae pv. tomato str. DC3000, and single members in several other Proteobacteria. The function is unknown.
The assembly of a macromolecular structure proceeds via a specific pathway of ordered events and occurs by changing of protein conformations as they join the assembly. The assembly process is aided by scaffolding proteins, which act as chaperones. In bacteriophages, scaffolding proteins B and D are responsible for procapsid formation. Copies of protein D (240) form the external scaffold, while 60 copies of protein B form the internal scaffold [
]. The structure of the scaffolding protein gpD is composed of six helices where one central helix is surrounded by 5 others.
Members of this uncharacterised protein family are found in Streptomyces, Anabaena sp. (strain PCC 7120), Clostridium acetobutylicum, Lactobacillus johnsonii NCC 533, Deinococcus radiodurans, and Pirellula sp. for a broad but sparse phylogenetic distribution that at least suggests lateral gene transfer.
Members of this family are small proteins, typically 73 amino acids in length, with single copies in each of several Proteobacteria, including Xylella fastidiosa, Pseudomonas aeruginosa, and Xanthomonas campestris. The function is unknown.
Som1 is a component of the mitochondrial protein export system. Som1 proteins exhibit a highly conserved region and a pattern of cysteine residues [
]. Stabilisation of Som1 occurs through an interaction between Som1 and Imp1, a peptidase required for proteolytic processing of certain proteins during their transport across the mitochondrial membrane []. This suggests that Som1 represents a third subunit of the Imp1 peptidase complex [].
This superfamily represents the chromosomal protein MC1, which protects DNA against thermal denaturation and shapes DNA by binding to it [
,
]. Its global fold consists of a pseudo barrel with an extension of the β-sheet (beta4-beta5) forming an arm (LP5) [].
Members of this uncharacterised protein family are found in a number of alphaproteobacteria, including root nodule bacteria, Brucella suis, Caulobacter crescentus (Caulobacter vibrioides), and Rhodopseudomonas palustris. Conserved residues include two well-separated cysteines, suggesting a disulphide bond. The function is unknown.
The lethal(2)giant larvae protein (Lgl) of Drosophila is essential for polarised epithelia development, for cell polarity associated with asymmetric cell division of neuroblasts during development and for oocyte polarity formation. As a result of alternative splicing Lgl has two isoforms that differ in their C-terminal region, isoform p78 which has an essential role in control of cell proliferation and differentiation during development as well as in controlling membrane trafficking underlying axonal growth and acts as a tumour suppressor [
,
,
]; and isoform p127 which has an accessory function in cell proliferation control and differentiation during development. These proteins, and their homologues, contain conserved functional domains such as homo-oligomerization domains, a cluster of phosphorylation sites and at least two WD-40 repeats, which suggests that these proteins have closely related functions [
,
].
Members of this family are small proteins, about 70 residues in length, with a basic triplet near the N terminus and a probable metal-binding motif CPXCX(18)CXXC. Members are found in various proteobacteria.
This family consists of bacterial proteins encoded within an intervening sequence present within some 23S rRNA genes [
,
,
]. It folds into an anti-parallel four-helix bundle and forms homopentamers [].
Members of this family are bacterial hypothetical proteins, about 160 amino acids in length, found in various proteobacteria, including members of the genera Pseudomonas and Vibrio. The C-terminal region is poorly conserved and is not included in the model.
This family of uncharacterised proteins is found in bacteria. Proteins in this family are typically between 100 and 143 amino acids in length. The N-terminal region is the most conserved.
In mammals, the second stage of spermatogenesis is characterised by the conversion of nucleosomal
chromatin to the compact, nonnucleosomal and transcriptionally inactive form found in the sperm nucleus.This condensation is associated with a double-protein transition. The first transition corresponds to the
replacement of histones by several spermatid-specific proteins, also called transition proteins, which arethemselves replaced by protamines during the second transition. Nuclear transition protein 2 (TP2) is one
of those spermatid-specific proteins. TP2 is a basic, zinc-binding protein [] of 116 to 137amino-acid residues.
Structurally, TP2 consists of three distinct parts, a conserved serine-rich N-terminal
domain of about 25 residues, a variable central domain of 20 to 50 residues which contains cysteine residues,and a conserved C-terminal domain of about 70 residues rich in lysines and arginines.
The development of B and T cells depends on the rearrangement of variable (V),
diversity (D), and joining (J) gene segments to produce mature Ig and T cellreceptor coding regions. This rearrangement process, known as V(D)J
recombination is initiated by a complex consisting of multi-domain proteins RAG1 and RAG2. The RAG proteins catalyse DNA cleavage in the first phase of thereaction using a recombination signal sequence (RSS) that flanks V, D and J
segments [,
,
].Recombination activating protein 1 (RAG1) is the catalytic component of the RAG complex [
]. RAG1 contains a RING finger domain that can act as a ubiquitin ligase (E3), and can promote its own ubiquitylation and targets both karyopherin alpha 1 (KPNA1) and histone 3 (H3) [,
].Many of the proteins recognised by this entry are fragments.
This entry consists of a relatively rare prokaryotic protein family (about 8 occurrences per 200 genomes). Genes for members of this family appear to be associated variously with phage and plasmid regions, restriction system loci, transposons, and housekeeping genes. Their function is unknown.
AcuA is part of the acuABC operon, which is possibly involved in the breakdown of acetoin and butanediol [
]. It is also a acetyltransferase that controls the activity of the acetyl coenzyme a synthetase (AcsA) in Bacillus subtilis [,
]. It has a role in cell growth and sporulation on acetoin or butanediol [], and may be involved in the breakdown of these compounds as a source of carbon.
Antifreeze proteins (AFPs) are a class of proteins that are able to bind to and inhibit the growth of macromolecular ice, thereby permitting an organism to survive subzero temperatures by decreasing the probability of ice nucleation in their bodies [
]. These proteins have been characterised from a variety of organisms, including fish, plants, bacteria, fungi and arthropods. This entry represents insect AFPs of the type found in spruce budworm, Choristoneura fumiferana.The structure of these AFPs consists of a left-handed β-helix with 15 residues per coil [
]. The β-helices of insect AFPs present a highly rigid array of threonine residues and bound water molecules that can effectively mimic the ice lattice. As such, β-helical AFPs provide a more effective coverage of the ice surface compared to the α-helical fish AFPs.A second insect antifreeze from Tenebrio molitor (
) also consists of β-helices, however in these proteins the helices form a right-handed twist; these proteins show no sequence homology to the current entry, but may act by a similar mechanism. The β-helix motif may be used as an AFP structural motif in non-homologous proteins from other (non-fish) organisms as well.
This family of proteins is found in viruses. Proteins in this family are typically between 241 and 261 amino acids in length. These proteins are capsid proteins from various astrovirus strains.
This entry represents a domain found in the tail assembly protein G from lambda-like viruses and their prophage.In bacteriophage lambda, the overlapping open reading frames G and T are expressed by a programmed translational frameshift to produce the tail assembly proteins G and GT [
]. Tail assembly protein GT shares it's N-terminal residues with tail assembly protein G, followed by residues of unique sequence []. An analogous frameshift is widely conserved among other dsDNA tailed phages in their corresponding 'G' and 'GT' tail genes even in the absence of detectable sequence homology []. The lambda tail assembly protein G and frameshift product GT are produced in a molar ratio of approximately 30:1[]. The correct molar ratio of these two related proteins, normally determined by the efficiency of the frameshift, is crucial for efficient assembly of functional tails []. Although tail assembly proteins G and GT are both required for assembly of functional tails, neither is present in mature tails [].
This domain is found in eukaryotes, and is approximately 60 amino acids in length. There is a single completely conserved residue C that may be functionally important. This is a calcium binding domain from the fungal protein CBP (calcium binding protein). This protein is a virulence factor with unknown virulence mechanisms. CBP complexes as a highly intertwined homodimer. Each monomer is comprised of four alpha helices which adopt the saposin fold, characteristic of a protein family that binds to membranes and lipids.
This entry represents an uncharacterised, well-conserved family of proteins found in bacteriophage and prophage regions of mainly Gram-positive bacteria.
This family of proteins is found in eukaryotes. Proteins in this family are typically between 165 and 237 amino acids in length. CD99L2 and CD99 are involved in trans-endothelial migration of neutrophils in vitro and in the recruitment of neutrophils into inflamed peritoneum.
This family of proteins is found in eukaryotes. Proteins in this family are typically between 285 and 331 amino acids in length. BSMAP has a putative transmembrane domain and is predicted to be a type I membrane glycoprotein.
This domain represents the N-terminal region of HoxA13 and other Hox proteins (HoxB13, HoxC13 and HoxD13). HoxA13 is involved in formation of the digital arch of the hands and feet, as well as in correct genital formation [
]. This domain is found in association with .
