Identification of a Distinct Substrate-binding Domain in the Bacterial Cysteine Methyltransferase Effectors NleE and OspZ.

2.50
Hdl Handle:
http://hdl.handle.net/10033/621274
Title:
Identification of a Distinct Substrate-binding Domain in the Bacterial Cysteine Methyltransferase Effectors NleE and OspZ.
Authors:
Zhang, Ying; Mühlen, Sabrina; Oates, Clare V; Pearson, Jaclyn S; Hartland, Elizabeth L
Abstract:
The type III secretion system effector protein NleE from enteropathogenic Escherichia coli plays a key role in the inhibition of NF-κB activation during infection. NleE inactivates the ubiquitin chain binding activity of host proteins TAK1-binding proteins 2 and 3 (TAB2 and TAB3) by modifying the Npl4 zinc finger domain through S-adenosyl methionine-dependent cysteine methylation. Using yeast two-hybrid protein interaction studies, we found that a conserved region between amino acids 34 and 52 of NleE, in particular the motif (49)GITR(52), was critical for TAB2 and TAB3 binding. NleE mutants lacking (49)GITR(52) were unable to methylate TAB3, and wild type NleE but not NleE(49AAAA52) where each of GITR was replaced with alanine restored the ability of an nleE mutant to inhibit IL-8 production during infection. Another NleE target, ZRANB3, also associated with NleE through the (49)GITR(52) motif. Ectopic expression of an N-terminal fragment of NleE (NleE(34-52)) in HeLa cells showed competitive inhibition of wild type NleE in the suppression of IL-8 secretion during enteropathogenic E. coli infection. Similar results were observed for the NleE homologue OspZ from Shigella flexneri 6 that also bound TAB3 through the (49)GITR(52) motif and decreased IL-8 transcription through modification of TAB3. In summary, we have identified a unique substrate-binding motif in NleE and OspZ that is required for the ability to inhibit the host inflammatory response.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Identification of a Distinct Substrate-binding Domain in the Bacterial Cysteine Methyltransferase Effectors NleE and OspZ. 2016, 291 (38):20149-62 J. Biol. Chem.
Journal:
The Journal of biological chemistry
Issue Date:
2016
URI:
http://hdl.handle.net/10033/621274
DOI:
10.1074/jbc.M116.734079
PubMed ID:
27445336
Type:
Article
Language:
en
ISSN:
1083-351X
Appears in Collections:
publications of the department of molecular Infectionbiology (MIBI)

Full metadata record

DC FieldValue Language
dc.contributor.authorZhang, Yingen
dc.contributor.authorMühlen, Sabrinaen
dc.contributor.authorOates, Clare Ven
dc.contributor.authorPearson, Jaclyn Sen
dc.contributor.authorHartland, Elizabeth Len
dc.date.accessioned2018-02-09T15:01:31Z-
dc.date.available2018-02-09T15:01:31Z-
dc.date.issued2016-
dc.identifier.citationIdentification of a Distinct Substrate-binding Domain in the Bacterial Cysteine Methyltransferase Effectors NleE and OspZ. 2016, 291 (38):20149-62 J. Biol. Chem.en
dc.identifier.issn1083-351X-
dc.identifier.pmid27445336-
dc.identifier.doi10.1074/jbc.M116.734079-
dc.identifier.urihttp://hdl.handle.net/10033/621274-
dc.description.abstractThe type III secretion system effector protein NleE from enteropathogenic Escherichia coli plays a key role in the inhibition of NF-κB activation during infection. NleE inactivates the ubiquitin chain binding activity of host proteins TAK1-binding proteins 2 and 3 (TAB2 and TAB3) by modifying the Npl4 zinc finger domain through S-adenosyl methionine-dependent cysteine methylation. Using yeast two-hybrid protein interaction studies, we found that a conserved region between amino acids 34 and 52 of NleE, in particular the motif (49)GITR(52), was critical for TAB2 and TAB3 binding. NleE mutants lacking (49)GITR(52) were unable to methylate TAB3, and wild type NleE but not NleE(49AAAA52) where each of GITR was replaced with alanine restored the ability of an nleE mutant to inhibit IL-8 production during infection. Another NleE target, ZRANB3, also associated with NleE through the (49)GITR(52) motif. Ectopic expression of an N-terminal fragment of NleE (NleE(34-52)) in HeLa cells showed competitive inhibition of wild type NleE in the suppression of IL-8 secretion during enteropathogenic E. coli infection. Similar results were observed for the NleE homologue OspZ from Shigella flexneri 6 that also bound TAB3 through the (49)GITR(52) motif and decreased IL-8 transcription through modification of TAB3. In summary, we have identified a unique substrate-binding motif in NleE and OspZ that is required for the ability to inhibit the host inflammatory response.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subject.meshAdaptor Proteins, Signal Transducingen
dc.subject.meshAmino Acid Motifsen
dc.subject.meshDNA Helicasesen
dc.subject.meshDysentery, Bacillaryen
dc.subject.meshEnteropathogenic Escherichia colien
dc.subject.meshEscherichia coli Infectionsen
dc.subject.meshEscherichia coli Proteinsen
dc.subject.meshHeLa Cellsen
dc.subject.meshHumansen
dc.subject.meshIntracellular Signaling Peptides and Proteinsen
dc.subject.meshProtein Bindingen
dc.subject.meshShigella flexnerien
dc.subject.meshVirulence Factorsen
dc.titleIdentification of a Distinct Substrate-binding Domain in the Bacterial Cysteine Methyltransferase Effectors NleE and OspZ.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalThe Journal of biological chemistryen
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