NLEB_CITRO
ID NLEB_CITRO Reviewed; 329 AA.
AC A0A482PDI9; Q5XMK8;
DT 02-JUN-2021, integrated into UniProtKB/Swiss-Prot.
DT 05-JUN-2019, sequence version 1.
DT 03-AUG-2022, entry version 8.
DE RecName: Full=Protein-arginine N-acetylglucosaminyltransferase NleB {ECO:0000305};
DE Short=Arginine GlcNAcyltransferase NleB {ECO:0000305};
DE Short=NleBc {ECO:0000303|PubMed:30979585};
DE EC=2.4.1.- {ECO:0000269|PubMed:23955153, ECO:0000269|PubMed:28860194, ECO:0000269|PubMed:31974499};
DE AltName: Full=Non-LEE-encoded type III effector B {ECO:0000303|PubMed:14988506, ECO:0000303|PubMed:16552063};
GN Name=nleB {ECO:0000303|PubMed:14988506, ECO:0000303|PubMed:16552063};
GN ORFNames=E2R62_01920 {ECO:0000312|EMBL:QBY27713.1};
OS Citrobacter rodentium.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Citrobacter.
OX NCBI_TaxID=67825;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=DBS100;
RX PubMed=14988506; DOI=10.1073/pnas.0400326101;
RA Deng W., Puente J.L., Gruenheid S., Li Y., Vallance B.A., Vazquez A.,
RA Barba J., Ibarra J.A., O'Donnell P., Metalnikov P., Ashman K., Lee S.,
RA Goode D., Pawson T., Finlay B.B.;
RT "Dissecting virulence: systematic and functional analyses of a
RT pathogenicity island.";
RL Proc. Natl. Acad. Sci. U.S.A. 101:3597-3602(2004).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DBS100;
RA Popov G., Fiebig A., Shideler S., Coombes B., Savchenko A.;
RT "Complete genome sequence of enteropathogenic Citrobacter rodentium strain
RT DBS100.";
RL Submitted (MAR-2019) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP FUNCTION, AND SUBCELLULAR LOCATION.
RX PubMed=16552063; DOI=10.1128/iai.74.4.2328-2337.2006;
RA Kelly M., Hart E., Mundy R., Marches O., Wiles S., Badea L., Luck S.,
RA Tauschek M., Frankel G., Robins-Browne R.M., Hartland E.L.;
RT "Essential role of the type III secretion system effector NleB in
RT colonization of mice by Citrobacter rodentium.";
RL Infect. Immun. 74:2328-2337(2006).
RN [4]
RP FUNCTION, AND MUTAGENESIS OF 221-ASP--ASP-223.
RX PubMed=23332158; DOI=10.1016/j.chom.2012.11.010;
RA Gao X., Wang X., Pham T.H., Feuerbacher L.A., Lubos M.L., Huang M.,
RA Olsen R., Mushegian A., Slawson C., Hardwidge P.R.;
RT "NleB, a bacterial effector with glycosyltransferase activity, targets
RT GAPDH function to inhibit NF-kappaB activation.";
RL Cell Host Microbe 13:87-99(2013).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, AND DISRUPTION PHENOTYPE.
RX PubMed=23955153; DOI=10.1038/nature12436;
RA Li S., Zhang L., Yao Q., Li L., Dong N., Rong J., Gao W., Ding X., Sun L.,
RA Chen X., Chen S., Shao F.;
RT "Pathogen blocks host death receptor signalling by arginine GlcNAcylation
RT of death domains.";
RL Nature 501:242-246(2013).
RN [6]
RP FUNCTION.
RX PubMed=24025841; DOI=10.1038/nature12524;
RA Pearson J.S., Giogha C., Ong S.Y., Kennedy C.L., Kelly M., Robinson K.S.,
RA Lung T.W., Mansell A., Riedmaier P., Oates C.V., Zaid A., Muehlen S.,
RA Crepin V.F., Marches O., Ang C.S., Williamson N.A., O'Reilly L.A.,
RA Bankovacki A., Nachbur U., Infusini G., Webb A.I., Silke J., Strasser A.,
RA Frankel G., Hartland E.L.;
RT "A type III effector antagonizes death receptor signalling during bacterial
RT gut infection.";
RL Nature 501:247-251(2013).
RN [7]
RP MUTAGENESIS OF TYR-219 AND GLU-253.
RX PubMed=26883593; DOI=10.1128/iai.01523-15;
RA Wong Fok Lung T., Giogha C., Creuzburg K., Ong S.Y., Pollock G.L.,
RA Zhang Y., Fung K.Y., Pearson J.S., Hartland E.L.;
RT "Mutagenesis and functional analysis of the bacterial arginine
RT glycosyltransferase effector NleB1 from enteropathogenic Escherichia
RT coli.";
RL Infect. Immun. 84:1346-1360(2016).
RN [8]
RP FUNCTION, AND MUTAGENESIS OF 221-ASP--ASP-223.
RX PubMed=27387501; DOI=10.1074/jbc.m116.738278;
RA Gao X., Pham T.H., Feuerbacher L.A., Chen K., Hays M.P., Singh G.,
RA Rueter C., Hurtado-Guerrero R., Hardwidge P.R.;
RT "Citrobacter rodentium NleB protein inhibits tumor necrosis factor (TNF)
RT receptor-associated factor 3 (TRAF3) ubiquitination to reduce host type I
RT interferon production.";
RL J. Biol. Chem. 291:18232-18238(2016).
RN [9]
RP FUNCTION.
RX PubMed=28522607; DOI=10.1074/jbc.m117.790675;
RA El Qaidi S., Chen K., Halim A., Siukstaite L., Rueter C.,
RA Hurtado-Guerrero R., Clausen H., Hardwidge P.R.;
RT "NleB/SseK effectors from Citrobacter rodentium, Escherichia coli, and
RT Salmonella enterica display distinct differences in host substrate
RT specificity.";
RL J. Biol. Chem. 292:11423-11430(2017).
RN [10]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=28860194; DOI=10.1074/jbc.m117.805036;
RA Scott N.E., Giogha C., Pollock G.L., Kennedy C.L., Webb A.I.,
RA Williamson N.A., Pearson J.S., Hartland E.L.;
RT "The bacterial arginine glycosyltransferase effector NleB preferentially
RT modifies Fas-associated death domain protein (FADD).";
RL J. Biol. Chem. 292:17337-17350(2017).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF 221-ASP--ASP-223.
RX PubMed=31974499; DOI=10.1038/s41598-020-58062-y;
RA El Qaidi S., Scott N.E., Hays M.P., Geisbrecht B.V., Watkins S.,
RA Hardwidge P.R.;
RT "An intra-bacterial activity for a T3SS effector.";
RL Sci. Rep. 10:1073-1073(2020).
RN [12]
RP MUTAGENESIS OF TRP-49; 186-ASP--ARG-189; ASP-186; TYR-219; GLU-253 AND
RP TYR-283.
RX PubMed=30979585; DOI=10.1016/j.molcel.2019.03.028;
RA Ding J., Pan X., Du L., Yao Q., Xue J., Yao H., Wang D.C., Li S., Shao F.;
RT "Structural and functional insights into host death domains inactivation by
RT the bacterial arginine GlcNAcyltransferase effector.";
RL Mol. Cell 74:922-935(2019).
CC -!- FUNCTION: Protein-arginine N-acetylglucosaminyltransferase effector
CC that disrupts TNF signaling in infected cells, including NF-kappa-B
CC signaling, apoptosis and necroptosis (PubMed:16552063, PubMed:23955153,
CC PubMed:24025841, PubMed:28522607). Acts by catalyzing the transfer of a
CC single N-acetylglucosamine (GlcNAc) to a conserved arginine residue in
CC the death domain of host proteins FADD, TNFRSF1A and RIPK1: arginine
CC GlcNAcylation prevents homotypic/heterotypic death domain interactions
CC and assembly of the oligomeric TNF-alpha receptor complex, thereby
CC disrupting TNF signaling (PubMed:23955153, PubMed:28522607,
CC PubMed:28860194). Has preference for host FADD as substrate compared to
CC TNFRSF1A and RIPK1 (PubMed:28860194). Also acts on host proteins
CC without a death domain: catalyzes GlcNAcylation of host GAPDH protein,
CC thereby preventing GAPDH interaction with TRAF2 and TRAF3, leading to
CC inhibit NF-kappa-B signaling and type I interferon production,
CC respectively (PubMed:23332158, PubMed:27387501, PubMed:28522607). Also
CC displays intra-bacterial activity by mediating GlcNAcylation of
CC glutathione synthetase GshB (PubMed:31974499). Catalyzes auto-
CC GlcNAcylation, which is required for activity toward death domain-
CC containing host target proteins (By similarity).
CC {ECO:0000250|UniProtKB:B7UI21, ECO:0000269|PubMed:16552063,
CC ECO:0000269|PubMed:23332158, ECO:0000269|PubMed:23955153,
CC ECO:0000269|PubMed:24025841, ECO:0000269|PubMed:27387501,
CC ECO:0000269|PubMed:28522607, ECO:0000269|PubMed:28860194,
CC ECO:0000269|PubMed:31974499}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-arginyl-[protein] + UDP-N-acetyl-alpha-D-glucosamine = H(+)
CC + N(omega)-(N-acetyl-beta-D-glucosaminyl)-L-arginyl-[protein] + UDP;
CC Xref=Rhea:RHEA:66632, Rhea:RHEA-COMP:10532, Rhea:RHEA-COMP:17079,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29965, ChEBI:CHEBI:57705,
CC ChEBI:CHEBI:58223, ChEBI:CHEBI:167322;
CC Evidence={ECO:0000269|PubMed:23955153, ECO:0000269|PubMed:28860194,
CC ECO:0000269|PubMed:31974499};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66633;
CC Evidence={ECO:0000269|PubMed:23955153, ECO:0000269|PubMed:28860194,
CC ECO:0000269|PubMed:31974499};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000250|UniProtKB:B7UI21};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:16552063}. Host cell
CC {ECO:0000269|PubMed:16552063}. Note=Secreted via the type III secretion
CC system (TTSS). {ECO:0000269|PubMed:16552063}.
CC -!- DOMAIN: Adopts a GT-A fold and acts as an inverting enzyme that
CC converts the alpha-configuration in the UDP-N-acetyl-alpha-D-
CC glucosamine donor to the beta configuration in the N-linked (GlcNAc)
CC arginine product. {ECO:0000250|UniProtKB:B7UI21}.
CC -!- PTM: Auto-glycosylated: arginine GlcNAcylation is required for activity
CC toward death domain-containing host target proteins.
CC {ECO:0000250|UniProtKB:B7UI21}.
CC -!- DISRUPTION PHENOTYPE: Reduced colonization of mice in single infections
CC and reduced colonic hyperplasia. {ECO:0000269|PubMed:16552063,
CC ECO:0000269|PubMed:23955153}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase NleB family.
CC {ECO:0000305}.
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DR EMBL; AY747106; AAU95470.1; -; Genomic_DNA.
DR EMBL; CP038008; QBY27713.1; -; Genomic_DNA.
DR RefSeq; WP_012905389.1; NZ_JXUN01000210.1.
DR AlphaFoldDB; A0A482PDI9; -.
DR SMR; A0A482PDI9; -.
DR OMA; LHNYNAF; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0043657; C:host cell; IDA:UniProtKB.
DR GO; GO:0016757; F:glycosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0106362; F:protein-arginine N-acetylglucosaminyltransferase activity; IDA:UniProtKB.
DR GO; GO:0090729; F:toxin activity; IDA:UniProtKB.
DR GO; GO:0052031; P:modulation by symbiont of host defense response; IDA:UniProtKB.
DR GO; GO:0034053; P:modulation by symbiont of host defense-related programmed cell death; IDA:UniProtKB.
DR GO; GO:0085034; P:suppression by symbiont of host I-kappaB kinase/NF-kappaB cascade; IDA:UniProtKB.
PE 1: Evidence at protein level;
KW Glycoprotein; Glycosyltransferase; Manganese; Metal-binding; Secreted;
KW Toxin; Transferase; Virulence.
FT CHAIN 1..329
FT /note="Protein-arginine N-acetylglucosaminyltransferase
FT NleB"
FT /id="PRO_0000452590"
FT MOTIF 221..223
FT /note="DXD motif"
FT /evidence="ECO:0000305"
FT ACT_SITE 253
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 48..50
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 72
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 219..222
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 223
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 320
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 322
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 322
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 327..329
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 13
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 53
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 159
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 293
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT MUTAGEN 49
FT /note="W->A: Reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:30979585"
FT MUTAGEN 186..189
FT /note="DFFR->AFFA: Reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:30979585"
FT MUTAGEN 186
FT /note="D->A: Strongly reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:30979585"
FT MUTAGEN 219
FT /note="Y->A: Strongly reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:26883593,
FT ECO:0000269|PubMed:30979585"
FT MUTAGEN 221..223
FT /note="DAD->AAA: Abolished N-acetylglucosaminyltransferase
FT activity and ability to disrupt host NF-kappa-B signaling
FT and type I interferon production."
FT /evidence="ECO:0000269|PubMed:23332158,
FT ECO:0000269|PubMed:27387501, ECO:0000269|PubMed:31974499"
FT MUTAGEN 253
FT /note="E->A: Reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:26883593,
FT ECO:0000269|PubMed:30979585"
FT MUTAGEN 283
FT /note="Y->A: Strongly reduced ability to promote bacterial
FT colonization in mice."
FT /evidence="ECO:0000269|PubMed:30979585"
FT CONFLICT 248
FT /note="Missing (in Ref. 1; AAU95470)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 329 AA; 37562 MW; D922A44EB91D9E92 CRC64;
MLSPLNVLQF NFRGETALSD SAPLQTVSFA GKDYSMEPID EKTPILFQWF EARPERYGKG
EVPILNTKEH PYLSNIINAA KIENERVIGV LVDGDFTYEQ RKEFLSLEDE HQNIKIIYRE
NVDFSMYDKK LSDIYLENIH EQESYPASER DNYLLGLLRE ELKNIPYGKD SLIESYAEKR
GHTWFDFFRN LAVLKGGGLF TETGKTGCHN ISPCGGCIYL DADMIITDKL GVLYAPDGIA
VYVDCNDNRK SLENGAIVVN RSNHPALLAG LDIMKSKVDA HPYYDGVGKG LKRHFNYSSL
QDYNVFCNFI EFKHKNIIPN TSMYTNSSW
