ID GPI3_YEAST Reviewed; 452 AA.
AC P32363; D6W3J3; Q08918;
DT 01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT 16-JUN-2009, sequence version 4.
DT 27-MAR-2024, entry version 181.
DE RecName: Full=Phosphatidylinositol N-acetylglucosaminyltransferase GPI3 subunit;
DE Short=GPI-GlcNAc transferase complex subunit GPI3;
DE Short=GPI-GnT subunit GPI3;
DE EC=2.4.1.198 {ECO:0000305|PubMed:10970797};
DE AltName: Full=GlcNAc-PI synthesis protein;
GN Name=SPT14; Synonyms=CWH6, GPI3; OrderedLocusNames=YPL175W; ORFNames=P2269;
OS Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Saccharomycetaceae; Saccharomyces.
OX NCBI_TaxID=559292;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 204508 / S288c;
RX PubMed=1660567; DOI=10.1007/bf00290682;
RA Fassler J.S., Gray W., Lee J.P., Yu G., Gingerich G.;
RT "The Saccharomyces cerevisiae SPT14 gene is essential for normal expression
RT of the yeast transposon, Ty, as well as for expression of the HIS4 gene and
RT several genes in the mating pathway.";
RL Mol. Gen. Genet. 230:310-320(1991).
RN [2]
RP SEQUENCE REVISION.
RA Fassler J.S.;
RL Submitted (JAN-1992) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 204508 / S288c;
RX PubMed=9169875;
RA Bussey H., Storms R.K., Ahmed A., Albermann K., Allen E., Ansorge W.,
RA Araujo R., Aparicio A., Barrell B.G., Badcock K., Benes V., Botstein D.,
RA Bowman S., Brueckner M., Carpenter J., Cherry J.M., Chung E.,
RA Churcher C.M., Coster F., Davis K., Davis R.W., Dietrich F.S., Delius H.,
RA DiPaolo T., Dubois E., Duesterhoeft A., Duncan M., Floeth M., Fortin N.,
RA Friesen J.D., Fritz C., Goffeau A., Hall J., Hebling U., Heumann K.,
RA Hilbert H., Hillier L.W., Hunicke-Smith S., Hyman R.W., Johnston M.,
RA Kalman S., Kleine K., Komp C., Kurdi O., Lashkari D., Lew H., Lin A.,
RA Lin D., Louis E.J., Marathe R., Messenguy F., Mewes H.-W., Mirtipati S.,
RA Moestl D., Mueller-Auer S., Namath A., Nentwich U., Oefner P., Pearson D.,
RA Petel F.X., Pohl T.M., Purnelle B., Rajandream M.A., Rechmann S.,
RA Rieger M., Riles L., Roberts D., Schaefer M., Scharfe M., Scherens B.,
RA Schramm S., Schroeder M., Sdicu A.-M., Tettelin H., Urrestarazu L.A.,
RA Ushinsky S., Vierendeels F., Vissers S., Voss H., Walsh S.V., Wambutt R.,
RA Wang Y., Wedler E., Wedler H., Winnett E., Zhong W.-W., Zollner A.,
RA Vo D.H., Hani J.;
RT "The nucleotide sequence of Saccharomyces cerevisiae chromosome XVI.";
RL Nature 387:103-105(1997).
RN [4]
RP GENOME REANNOTATION.
RC STRAIN=ATCC 204508 / S288c;
RX PubMed=24374639; DOI=10.1534/g3.113.008995;
RA Engel S.R., Dietrich F.S., Fisk D.G., Binkley G., Balakrishnan R.,
RA Costanzo M.C., Dwight S.S., Hitz B.C., Karra K., Nash R.S., Weng S.,
RA Wong E.D., Lloyd P., Skrzypek M.S., Miyasato S.R., Simison M., Cherry J.M.;
RT "The reference genome sequence of Saccharomyces cerevisiae: Then and now.";
RL G3 (Bethesda) 4:389-398(2014).
RN [5]
RP CHARACTERIZATION.
RX PubMed=7737116; DOI=10.1002/j.1460-2075.1995.tb07152.x;
RA Schoenbaechler M., Horvath A., Fassler J.S., Riezman H.;
RT "The yeast spt14 gene is homologous to the human PIG-A gene and is required
RT for GPI anchor synthesis.";
RL EMBO J. 14:1637-1645(1995).
RN [6]
RP FUNCTION.
RX PubMed=7768896; DOI=10.1074/jbc.270.22.13029;
RA Leidich S.D., Kostova Z., Latek R.R., Costello L.C., Drapp D.A., Gray W.,
RA Fassler J.S., Orlean P.;
RT "Temperature-sensitive yeast GPI anchoring mutants gpi2 and gpi3 are
RT defective in the synthesis of N-acetylglucosaminyl phosphatidylinositol.
RT Cloning of the GPI2 gene.";
RL J. Biol. Chem. 270:13029-13035(1995).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND SUBSTRATE BINDING.
RX PubMed=10970797; DOI=10.1042/bj3500815;
RA Kostova Z., Rancour D.M., Menon A.K., Orlean P.;
RT "Photoaffinity labelling with P3-(4-azidoanilido)uridine 5'-triphosphate
RT identifies gpi3p as the UDP-GlcNAc-binding subunit of the enzyme that
RT catalyses formation of GlcNAc-phosphatidylinositol, the first glycolipid
RT intermediate in glycosylphosphatidylinositol synthesis.";
RL Biochem. J. 350:815-822(2000).
RN [8]
RP IDENTIFICATION OF INTRON.
RX PubMed=10734188; DOI=10.1093/nar/28.8.1700;
RA Davis C.A., Grate L., Spingola M., Ares M. Jr.;
RT "Test of intron predictions reveals novel splice sites, alternatively
RT spliced mRNAs and new introns in meiotically regulated genes of yeast.";
RL Nucleic Acids Res. 28:1700-1706(2000).
RN [9]
RP MUTAGENESIS OF GLU-289; GLU-297 AND CYS-301.
RX PubMed=14622279; DOI=10.1046/j.1432-1033.2003.03844.x;
RA Kostova Z., Yan B.C., Vainauskas S., Schwartz R., Menon A.K., Orlean P.;
RT "Comparative importance in vivo of conserved glutamate residues in the EX7E
RT motif retaining glycosyltransferase Gpi3p, the UDP-GlcNAc-binding subunit
RT of the first enzyme in glycosylphosphatidylinositol assembly.";
RL Eur. J. Biochem. 270:4507-4514(2003).
RN [10]
RP LEVEL OF PROTEIN EXPRESSION [LARGE SCALE ANALYSIS].
RX PubMed=14562106; DOI=10.1038/nature02046;
RA Ghaemmaghami S., Huh W.-K., Bower K., Howson R.W., Belle A., Dephoure N.,
RA O'Shea E.K., Weissman J.S.;
RT "Global analysis of protein expression in yeast.";
RL Nature 425:737-741(2003).
RN [11]
RP ACTIVITY REGULATION.
RX PubMed=15163411; DOI=10.1016/j.cell.2004.05.003;
RA Sobering A.K., Watanabe R., Romeo M.J., Yan B.C., Specht C.A., Orlean P.,
RA Riezman H., Levin D.E.;
RT "Yeast Ras regulates the complex that catalyzes the first step in GPI-
RT anchor biosynthesis at the ER.";
RL Cell 117:637-648(2004).
CC -!- FUNCTION: Catalytic subunit in the complex catalyzing the transfer of
CC N-acetylglucosamine from UDP-N-acetylglucosamine to
CC phosphatidylinositol, the first step of GPI biosynthesis.
CC {ECO:0000269|PubMed:10970797, ECO:0000269|PubMed:7768896}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 1,2-diacyl-sn-glycero-3-phospho-(1D-myo-inositol) + UDP-N-
CC acetyl-alpha-D-glucosamine = a 6-(N-acetyl-alpha-D-glucosaminyl)-1-
CC phosphatidyl-1D-myo-inositol + H(+) + UDP; Xref=Rhea:RHEA:14789,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57265, ChEBI:CHEBI:57705,
CC ChEBI:CHEBI:57880, ChEBI:CHEBI:58223; EC=2.4.1.198;
CC Evidence={ECO:0000305|PubMed:10970797};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:14790;
CC Evidence={ECO:0000305|PubMed:10970797};
CC -!- ACTIVITY REGULATION: Inhibited by Ras, probably via the interaction
CC between RAS2 and ERI1. {ECO:0000269|PubMed:15163411}.
CC -!- PATHWAY: Glycolipid biosynthesis; glycosylphosphatidylinositol-anchor
CC biosynthesis. {ECO:0000305|PubMed:7737116}.
CC -!- SUBUNIT: Component of the phosphatidylinositol N-
CC acetylglucosaminyltransferase (GPI-GlcNAc transferase) complex composed
CC of at least GPI1, GPI2, GPI3, GPI15, GPI19 and ERI1.
CC {ECO:0000305|PubMed:10970797}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane {ECO:0000305};
CC Single-pass membrane protein {ECO:0000305}.
CC -!- MISCELLANEOUS: Present with 1480 molecules/cell in log phase SD medium.
CC {ECO:0000269|PubMed:14562106}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase group 1 family.
CC Glycosyltransferase 4 subfamily. {ECO:0000305}.
CC -!- CAUTION: Was originally thought to be involved in transcription.
CC {ECO:0000305|PubMed:1660567}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAA97882.1; Type=Erroneous gene model prediction; Evidence={ECO:0000305};
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DR EMBL; X63290; CAA44924.1; -; Genomic_DNA.
DR EMBL; Z73531; CAA97882.1; ALT_SEQ; Genomic_DNA.
DR EMBL; BK006949; DAA11259.1; -; Genomic_DNA.
DR PIR; S65187; S65187.
DR RefSeq; NP_015150.2; NM_001183989.1.
DR AlphaFoldDB; P32363; -.
DR SMR; P32363; -.
DR BioGRID; 36008; 69.
DR ComplexPortal; CPX-1274; Glycosylphosphatidylinositol-N-acetylglucosaminyltransferase complex.
DR DIP; DIP-1834N; -.
DR IntAct; P32363; 2.
DR MINT; P32363; -.
DR STRING; 4932.YPL175W; -.
DR CAZy; GT4; Glycosyltransferase Family 4.
DR iPTMnet; P32363; -.
DR MaxQB; P32363; -.
DR PaxDb; 4932-YPL175W; -.
DR PeptideAtlas; P32363; -.
DR EnsemblFungi; YPL175W_mRNA; YPL175W; YPL175W.
DR GeneID; 855928; -.
DR KEGG; sce:YPL175W; -.
DR AGR; SGD:S000006096; -.
DR SGD; S000006096; SPT14.
DR VEuPathDB; FungiDB:YPL175W; -.
DR eggNOG; KOG1111; Eukaryota.
DR GeneTree; ENSGT00390000014405; -.
DR HOGENOM; CLU_009583_19_0_1; -.
DR InParanoid; P32363; -.
DR OMA; QCVLPTM; -.
DR OrthoDB; 24420at2759; -.
DR BioCyc; YEAST:G3O-34070-MONOMER; -.
DR UniPathway; UPA00196; -.
DR BioGRID-ORCS; 855928; 0 hits in 10 CRISPR screens.
DR PRO; PR:P32363; -.
DR Proteomes; UP000002311; Chromosome XVI.
DR RNAct; P32363; Protein.
DR GO; GO:0005829; C:cytosol; HDA:SGD.
DR GO; GO:0005783; C:endoplasmic reticulum; IMP:SGD.
DR GO; GO:0005789; C:endoplasmic reticulum membrane; NAS:ComplexPortal.
DR GO; GO:0000506; C:glycosylphosphatidylinositol-N-acetylglucosaminyltransferase (GPI-GnT) complex; IBA:GO_Central.
DR GO; GO:0017176; F:phosphatidylinositol N-acetylglucosaminyltransferase activity; IMP:SGD.
DR GO; GO:0031505; P:fungal-type cell wall organization; NAS:ComplexPortal.
DR GO; GO:0006506; P:GPI anchor biosynthetic process; IBA:GO_Central.
DR CDD; cd03796; GT4_PIG-A-like; 1.
DR Gene3D; 3.40.50.2000; Glycogen Phosphorylase B; 2.
DR InterPro; IPR001296; Glyco_trans_1.
DR InterPro; IPR039507; PIG-A/GPI3.
DR InterPro; IPR013234; PIGA_GPI_anchor_biosynthesis.
DR PANTHER; PTHR45871; N-ACETYLGLUCOSAMINYL-PHOSPHATIDYLINOSITOL BIOSYNTHETIC PROTEIN; 1.
DR PANTHER; PTHR45871:SF1; PHOSPHATIDYLINOSITOL N-ACETYLGLUCOSAMINYLTRANSFERASE SUBUNIT A; 1.
DR Pfam; PF00534; Glycos_transf_1; 1.
DR Pfam; PF08288; PIGA; 1.
DR SUPFAM; SSF53756; UDP-Glycosyltransferase/glycogen phosphorylase; 1.
PE 1: Evidence at protein level;
KW Endoplasmic reticulum; Glycosyltransferase; GPI-anchor biosynthesis;
KW Membrane; Reference proteome; Transferase; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..452
FT /note="Phosphatidylinositol N-acetylglucosaminyltransferase
FT GPI3 subunit"
FT /id="PRO_0000080325"
FT TRANSMEM 407..427
FT /note="Helical"
FT /evidence="ECO:0000255"
FT MUTAGEN 289
FT /note="E->A: Severe growth defect."
FT /evidence="ECO:0000269|PubMed:14622279"
FT MUTAGEN 289
FT /note="E->D: Reduces the transferase reaction 12-fold."
FT /evidence="ECO:0000269|PubMed:14622279"
FT MUTAGEN 289
FT /note="E->G: Lethal."
FT /evidence="ECO:0000269|PubMed:14622279"
FT MUTAGEN 297
FT /note="E->A,G: Lethal."
FT /evidence="ECO:0000269|PubMed:14622279"
FT MUTAGEN 297
FT /note="E->D: No transferase activity."
FT /evidence="ECO:0000269|PubMed:14622279"
FT MUTAGEN 301
FT /note="C->A: Reduces the transferase reaction 5-fold."
FT /evidence="ECO:0000269|PubMed:14622279"
SQ SEQUENCE 452 AA; 51242 MW; 7589FD815707EC73 CRC64;
MGFNIAMLCD FFYPQLGGVE FHIYHLSQKL IDLGHSVVII THAYKDRVGV RHLTNGLKVY
HVPFFVIFRE TTFPTVFSTF PIIRNILLRE QIQIVHSHGS ASTFAHEGIL HANTMGLRTV
FTDHSLYGFN NLTSIWVNKL LTFTLTNIDR VICVSNTCKE NMIVRTELSP DIISVIPNAV
VSEDFKPRDP TGGTKRKQSR DKIVIVVIGR LFPNKGSDLL TRIIPKVCSS HEDVEFIVAG
DGPKFIDFQQ MIESHRLQKR VQLLGSVPHE KVRDVLCQGD IYLHASLTEA FGTILVEAAS
CNLLIVTTQV GGIPEVLPNE MTVYAEQTSV SDLVQATNKA INIIRSKALD TSSFHDSVSK
MYDWMDVAKR TVEIYTNISS TSSADDKDWM KMVANLYKRD GIWAKHLYLL CGIVEYMLFF
LLEWLYPRDE IDLAPKWPKK TVSNETKEAR ET
//
