Home LiteratureArticle Details
PMID: 18845161 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structure of the cyclomodulin Cif from pathogenic Escherichia coli.

Journal of molecular biology ·Vol. 384 ·No. 2 ·2008-12-12 ·Pages 465-77

Hsu Y, Jubelin G, Taieb F, Nougayrède JP, Oswald E, Stebbins CE

Abstract

Bacterial pathogens have evolved a sophisticated arsenal of virulence factors to modulate host cell biology. Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC) use a type III protein secretion system (T3SS) to inject microbial proteins into host cells. The T3SS effector cycle inhibiting factor (Cif) produced by EPEC and EHEC is able to block host eukaryotic cell-cycle progression. We present here a crystal structure of Cif, revealing it to be a divergent member of the superfamily of enzymes including cysteine proteases and acetyltransferases that share a common catalytic triad. Mutation of these conserved active site residues abolishes the ability of Cif to block cell-cycle progression. Finally, we demonstrate that irreversible cysteine protease inhibitors do not abolish the Cif cytopathic effect, suggesting that another enzymatic activity may underlie the biological activity of this virulence factor.

MeSH Terms
Actins/metabolism Amino Acid Sequence Caseins/metabolism Catalysis/drug effects Cell Cycle/drug effects Chromatography, Gel Crystallography, X-Ray Cysteine Endopeptidases/chemistry Enzyme Activation/drug effects Escherichia coli/enzymology,pathogenicity Escherichia coli Proteins/antagonists & inhibitors,chemistry HeLa Cells Humans Models, Molecular Molecular Sequence Data Phenotype Protease Inhibitors/pharmacology Protein Structure, Secondary Sequence Deletion Stress Fibers/drug effects,enzymology Structural Homology, Protein Substrate Specificity/drug effects
Chemicals
Actins Caseins Cif protein, E coli Escherichia coli Proteins Protease Inhibitors Cysteine Endopeptidases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hsu Yun
Laboratory of Structural Microbiology, Rockefeller University, New York, NY 10021, USA.
Jubelin Gregory
Taieb Frédéric
Nougayrède Jean-Philippe
Oswald Eric
Stebbins C Erec
References (33)
33 references, click to expand
  1. Automated MAD and MIR structure solution.
    Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):849-61 PMID: 10089316
  2. The bacterial cytolethal distending toxin (CDT) triggers a G2 cell cycle checkpoint in mammalian cells without preliminary induction of DNA strand breaks.
    Oncogene. 1999 Nov 4;18(46):6296-304 PMID: 10597228
  3. Structure of arylamine N-acetyltransferase reveals a catalytic triad.
    Nat Struct Biol. 2000 Jul;7(7):560-4 PMID: 10876241
  4. A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein.
    Science. 2000 Oct 13;290(5490):354-7 PMID: 11030657
  5. Disruption of signaling by Yersinia effector YopJ, a ubiquitin-like protein protease.
    Science. 2000 Nov 24;290(5496):1594-7 PMID: 11090361
  6. Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor.
    Biochem J. 2000 Dec 15;352 Pt 3:739-45 PMID: 11104681
  7. Homology modelling and structural analysis of human arylamine N-acetyltransferase NAT1: evidence for the conservation of a cysteine protease catalytic domain and an active-site loop.
    Biochem J. 2001 Jun 1;356(Pt 2):327-34 PMID: 11368758
  8. Type III secretion-dependent cell cycle block caused in HeLa cells by enteropathogenic Escherichia coli O103.
    Infect Immun. 2001 Nov;69(11):6785-95 PMID: 11598051
  9. A Yersinia effector and a Pseudomonas avirulence protein define a family of cysteine proteases functioning in bacterial pathogenesis.
    Cell. 2002 May 31;109(5):575-88 PMID: 12062101
  10. Enteropathogenic and enterohaemorrhagic Escherichia coli deliver a novel effector called Cif, which blocks cell cycle G2/M transition.
    Mol Microbiol. 2003 Dec;50(5):1553-67 PMID: 14651638
  11. SCOP database in 2004: refinements integrate structure and sequence family data.
    Nucleic Acids Res. 2004 Jan 1;32(Database issue):D226-9 PMID: 14681400
  12. The crystal structure of Pseudomonas avirulence protein AvrPphB: a papain-like fold with a distinct substrate-binding site.
    Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):302-7 PMID: 14694194
  13. Macromolecular TLS refinement in REFMAC at moderate resolutions.
    Methods Enzymol. 2003;374:300-21 PMID: 14696379
  14. Assembly and function of a bacterial genotoxin.
    Nature. 2004 May 27;429(6990):429-33 PMID: 15164065
  15. Identification of the secretion and translocation domain of the enteropathogenic and enterohemorrhagic Escherichia coli effector Cif, using TEM-1 beta-lactamase as a new fluorescence-based reporter.
    J Bacteriol. 2004 Aug;186(16):5486-95 PMID: 15292151
  16. wARP: improvement and extension of crystallographic phases by weighted averaging of multiple-refined dummy atomic models.
    Acta Crystallogr D Biol Crystallogr. 1997 Jul 1;53(Pt 4):448-55 PMID: 15299911
  17. Refinement of macromolecular structures by the maximum-likelihood method.
    Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55 PMID: 15299926
  18. Host-pathogen interactions: a diversity of themes, a variety of molecular machines.
    Curr Opin Microbiol. 2005 Feb;8(1):1-3 PMID: 15694849
  19. Bacterial toxins that modulate host cell-cycle progression.
    Curr Opin Microbiol. 2005 Feb;8(1):83-91 PMID: 15694861
  20. Cyclomodulins: bacterial effectors that modulate the eukaryotic cell cycle.
    Trends Microbiol. 2005 Mar;13(3):103-10 PMID: 15737728
  21. Activation of a phytopathogenic bacterial effector protein by a eukaryotic cyclophilin.
    Science. 2005 Apr 22;308(5721):548-50 PMID: 15746386
  22. Enteropathogenic and enterohemorrhagic Escherichia coli infections: translocation, translocation, translocation.
    Infect Immun. 2005 May;73(5):2573-85 PMID: 15845459
  23. Yersinia YopJ acetylates and inhibits kinase activation by blocking phosphorylation.
    Science. 2006 May 26;312(5777):1211-4 PMID: 16728640
  24. Escherichia coli cyclomodulin Cif induces G2 arrest of the host cell cycle without activation of the DNA-damage checkpoint-signalling pathway.
    Cell Microbiol. 2006 Dec;8(12):1910-21 PMID: 16848790
  25. Escherichia coli induces DNA double-strand breaks in eukaryotic cells.
    Science. 2006 Aug 11;313(5788):848-51 PMID: 16902142
  26. Protein delivery into eukaryotic cells by type III secretion machines.
    Nature. 2006 Nov 30;444(7119):567-73 PMID: 17136086
  27. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  28. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  29. Structure of papain refined at 1.65 A resolution.
    J Mol Biol. 1984 Oct 25;179(2):233-56 PMID: 6502713
  30. Families of cysteine peptidases.
    Methods Enzymol. 1994;244:461-86 PMID: 7845226
  31. Dali: a network tool for protein structure comparison.
    Trends Biochem Sci. 1995 Nov;20(11):478-80 PMID: 8578593
  32. Structural determinants of the specificity for synaptic vesicle-associated membrane protein/synaptobrevin of tetanus and botulinum type B and G neurotoxins.
    J Biol Chem. 1996 Aug 23;271(34):20353-8 PMID: 8702770
  33. Enteroaggregative Escherichia coli.
    Emerg Infect Dis. 1998 Apr-Jun;4(2):251-61 PMID: 9621195
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
1089-8638
DOI
10.1016/j.jmb.2008.09.051
Published
2008-12-12
Epub
2008-00-27
Pages
465-77
Language
English
Region
England
NLM ID
2985088R
PMCID
PMC2659761
Subset
IM
Grants
NIAID NIH HHS · R01 AI052182-03 · United States
NIBIB NIH HHS · P30 EB009998 · United States
NIAID NIH HHS · R01 AI052182-04 · United States
NIAID NIH HHS · R01 AI052182-05 · United States
NIAID NIH HHS · R01 AI052182 · United States
Databases
PDB
3EFY
External Links
PubMed source DOI: 10.1016/j.jmb.2008.09.051 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2