GTF3_STRPA
ID GTF3_STRPA Reviewed; 330 AA.
AC B5A7L9;
DT 08-MAY-2019, integrated into UniProtKB/Swiss-Prot.
DT 23-SEP-2008, sequence version 1.
DT 03-AUG-2022, entry version 37.
DE RecName: Full=Glucosyltransferase 3 {ECO:0000255|HAMAP-Rule:MF_00841, ECO:0000303|PubMed:20164186};
DE EC=2.4.1.- {ECO:0000255|HAMAP-Rule:MF_00841, ECO:0000269|PubMed:20164186};
GN Name=gtf3 {ECO:0000255|HAMAP-Rule:MF_00841, ECO:0000303|PubMed:20164186};
GN Synonyms=nss {ECO:0000303|PubMed:17296746};
OS Streptococcus parasanguinis.
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=1318;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=FW213;
RX PubMed=17296746; DOI=10.1128/iai.01544-06;
RA Wu H., Zeng M., Fives-Taylor P.;
RT "The glycan moieties and the N-terminal polypeptide backbone of a fimbria-
RT associated adhesin, Fap1, play distinct roles in the biofilm development of
RT Streptococcus parasanguinis.";
RL Infect. Immun. 75:2181-2188(2007).
RN [2]
RP FUNCTION, PATHWAY, DOMAIN, AND DISRUPTION PHENOTYPE.
RC STRAIN=FW213;
RX PubMed=20164186; DOI=10.1074/jbc.m109.066928;
RA Zhou M., Zhu F., Dong S., Pritchard D.G., Wu H.;
RT "A novel glucosyltransferase is required for glycosylation of a serine-rich
RT adhesin and biofilm formation by Streptococcus parasanguinis.";
RL J. Biol. Chem. 285:12140-12148(2010).
RN [3]
RP FUNCTION, PATHWAY, AND MUTAGENESIS OF 106-MET--PHE-111.
RC STRAIN=FW213;
RX PubMed=25404702; DOI=10.1128/jb.02267-14;
RA Zhu F., Zhang H., Wu H.;
RT "A conserved domain is crucial for acceptor substrate binding in a family
RT of glucosyltransferases.";
RL J. Bacteriol. 197:510-517(2015).
RN [4] {ECO:0007744|PDB:3QKW, ECO:0007744|PDB:3RHZ}
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF 1-329 IN COMPLEX WITH UDP
RP PRODUCT, FUNCTION, COFACTOR, PATHWAY, SUBUNIT, DOMAIN, DISRUPTION
RP PHENOTYPE, AND MUTAGENESIS OF ARG-179; TYR-211; ASP-214; LYS-246; SER-249;
RP 314-PHE--ASP-330; PHE-314; PHE-315; ARG-318 AND LEU-320.
RC STRAIN=FW213;
RX PubMed=21653318; DOI=10.1074/jbc.m110.208629;
RA Zhu F., Erlandsen H., Ding L., Li J., Huang Y., Zhou M., Liang X., Ma J.,
RA Wu H.;
RT "Structural and functional analysis of a new subfamily of
RT glycosyltransferases required for glycosylation of serine-rich
RT streptococcal adhesins.";
RL J. Biol. Chem. 286:27048-27057(2011).
CC -!- FUNCTION: Required for polymorphic O-glycosylation of the serine-rich
CC repeat protein Fap1. Catalyzes the second step in glycosylation of the
CC serine-rich repeat protein in this bacteria. Transfers glucose from
CC UDP-glucose to the terminal GlcNAc moiety of 3-O-(N-acetyl-alpha-D-
CC glucosaminyl)-L-seryl-[protein] which results from the first
CC glycosylation step of Fap1; does not use other sugar nucleotides as
CC substrates. {ECO:0000269|PubMed:20164186, ECO:0000269|PubMed:21653318,
CC ECO:0000269|PubMed:25404702}.
CC -!- COFACTOR:
CC Note=In vitro glycosyltransferase activity is metal-independent.
CC {ECO:0000269|PubMed:21653318};
CC -!- PATHWAY: Protein modification; protein glycosylation.
CC {ECO:0000255|HAMAP-Rule:MF_00841, ECO:0000269|PubMed:20164186,
CC ECO:0000269|PubMed:25404702}.
CC -!- SUBUNIT: Homotetramer; a dimer of dimers. {ECO:0000255|HAMAP-
CC Rule:MF_00841, ECO:0000269|PubMed:21653318}.
CC -!- DOMAIN: Dimerizes via the C-terminus; dimerization is required for
CC tetramer formation (PubMed:21653318). Binds protein substrate via an
CC exposed loop in the N-terminus (PubMed:25404702).
CC {ECO:0000269|PubMed:21653318, ECO:0000269|PubMed:25404702}.
CC -!- DISRUPTION PHENOTYPE: The Fap1 protein no longer reacts with a glycan-
CC specific antibody, and migrates as a much larger than wild-type
CC protein. Fap1 is not able to undergo the second glycosylation step. Has
CC defects in biofilm mass accumulation, the biofilm that forms is not as
CC thick as wild-type (PubMed:20164186). Deletion of this gene can be
CC complemented by the ortholog from S.agalactiae strain COH1, but not
CC from S. pneumoniae strain TIGR4 or from S. sanguinis strain SK36
CC (PubMed:21653318). {ECO:0000269|PubMed:20164186,
CC ECO:0000269|PubMed:21653318}.
CC -!- SIMILARITY: Belongs to the Gtf3 glucosyltransferase family.
CC {ECO:0000255|HAMAP-Rule:MF_00841}.
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DR EMBL; EU821531; ACF35267.1; -; Genomic_DNA.
DR RefSeq; WP_014713999.1; NZ_JYPA01000014.1.
DR PDB; 3QKW; X-ray; 2.29 A; A/B/C/D=1-330.
DR PDB; 3RHZ; X-ray; 1.90 A; A/B=1-329.
DR PDBsum; 3QKW; -.
DR PDBsum; 3RHZ; -.
DR AlphaFoldDB; B5A7L9; -.
DR SMR; B5A7L9; -.
DR PATRIC; fig|1305.10.peg.1021; -.
DR UniPathway; UPA00378; -.
DR EvolutionaryTrace; B5A7L9; -.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0035251; F:UDP-glucosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0006486; P:protein glycosylation; IEA:UniProtKB-UniPathway.
DR HAMAP; MF_00841; Gtf3; 1.
DR InterPro; IPR043676; Gtf3.
PE 1: Evidence at protein level;
KW 3D-structure; Glycosyltransferase; Nucleotide-binding; Transferase.
FT CHAIN 1..330
FT /note="Glucosyltransferase 3"
FT /id="PRO_0000447236"
FT REGION 106..111
FT /note="Substrate protein-binding loop"
FT /evidence="ECO:0000269|PubMed:25404702"
FT BINDING 16
FT /ligand="UDP"
FT /ligand_id="ChEBI:CHEBI:58223"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00841,
FT ECO:0000269|PubMed:21653318, ECO:0007744|PDB:3QKW,
FT ECO:0007744|PDB:3RHZ"
FT BINDING 179
FT /ligand="UDP"
FT /ligand_id="ChEBI:CHEBI:58223"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00841,
FT ECO:0000269|PubMed:21653318, ECO:0007744|PDB:3QKW,
FT ECO:0007744|PDB:3RHZ"
FT BINDING 211..214
FT /ligand="UDP"
FT /ligand_id="ChEBI:CHEBI:58223"
FT /evidence="ECO:0000269|PubMed:21653318,
FT ECO:0007744|PDB:3QKW, ECO:0007744|PDB:3RHZ"
FT BINDING 244..249
FT /ligand="UDP"
FT /ligand_id="ChEBI:CHEBI:58223"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00841,
FT ECO:0000269|PubMed:21653318, ECO:0007744|PDB:3QKW,
FT ECO:0007744|PDB:3RHZ"
FT MUTAGEN 106..111
FT /note="Missing: Loss of binding to Fap1-GlcNAc substrate,
FT loss of glycosyltransferase activity."
FT /evidence="ECO:0000269|PubMed:25404702"
FT MUTAGEN 179
FT /note="R->A: Complete loss of glycosyltransferase activity,
FT does not restore Fap1 glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 211
FT /note="Y->A: 25% glycosyltransferase activity, partially
FT restores Fap1 glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 214
FT /note="D->A: Wild-type glycosyltransferase activity, fully
FT restores Fap1 glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 246
FT /note="K->A: Complete loss of glycosyltransferase activity,
FT the protein forms tetramers, does not restore Fap1
FT glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 249
FT /note="S->A: Wild-type glycosyltransferase activity."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 314..330
FT /note="Missing: Complete loss of glycosyltransferase
FT activity."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 314
FT /note="F->A: 25% glycosyltransferase activity, the protein
FT dimerizes but does not tetramerize."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 315
FT /note="F->A: Complete loss of glycosyltransferase activity,
FT the protein does not dimerize."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 318
FT /note="R->A: 25% glycosyltransferase activity, the protein
FT dimerizes but does not tetramerize, partially restores Fap1
FT glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT MUTAGEN 320
FT /note="L->A: Complete loss of glycosyltransferase activity,
FT the protein does not dimerize, does not restore Fap1
FT glycosylation in vivo."
FT /evidence="ECO:0000269|PubMed:21653318"
FT STRAND 3..11
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 16..30
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 34..39
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 43..45
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 48..58
FT /evidence="ECO:0007829|PDB:3RHZ"
FT TURN 59..61
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 67..72
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 78..88
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 94..100
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 103..106
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 108..113
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 114..121
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 125..130
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 132..140
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 146..150
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 166..173
FT /evidence="ECO:0007829|PDB:3RHZ"
FT TURN 177..179
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 181..185
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 192..197
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 206..210
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 214..222
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 224..228
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 233..235
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 236..239
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 245..253
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 257..260
FT /evidence="ECO:0007829|PDB:3RHZ"
FT TURN 264..266
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 267..272
FT /evidence="ECO:0007829|PDB:3RHZ"
FT STRAND 275..280
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 281..290
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 293..306
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 308..311
FT /evidence="ECO:0007829|PDB:3RHZ"
FT HELIX 314..328
FT /evidence="ECO:0007829|PDB:3RHZ"
SQ SEQUENCE 330 AA; 38053 MW; B307A8E48093F876 CRC64;
MRVYITNING QSIQSTAQLC QNTVTDVAVS LGYRELGIYC YQIHTDSESE LSKRLDGIVA
GLRHGDVVIF QTPTWNTTEF DEKLMNKLKL YDIKIVLFIH DVVPLMFSGN FYLMDRTIAY
YNKADVVVAP SQKMIDKLRD FGMNVSKTVV QGMWDHPTQA PMFPAGLKRE IHFPGNPERF
SFVKEWKYDI PLKVYTWQNV ELPQNVHKIN YRPDEQLLME MSQGGFGLVW MDDKDKEYQS
LYCSYKLGSF LAAGIPVIVQ EGIANQELIE NNGLGWIVKD VEEAIMKVKN VNEDEYIELV
KNVRSFNPIL RKGFFTRRLL TESVFQAICD
