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PMID: 9916051 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Deamidation of Cdc42 and Rac by Escherichia coli cytotoxic necrotizing factor 1: activation of c-Jun N-terminal kinase in HeLa cells.

Infection and immunity ·Vol. 67 ·No. 2 ·1999-02-00 ·Pages 496-503

Lerm M, Selzer J, Hoffmeyer A, Rapp UR, Aktories K, Schmidt G

Abstract

Recently, Escherichia coli cytotoxic necrotizing factor 1 (CNF1) was shown to activate the low-molecular-mass GTPase RhoA by deamidation of Gln63, thereby inhibiting intrinsic and GTPase-activating protein (GAP)-stimulated GTPase activities (G. Schmidt, P. Sehr, M. Wilm, J. Selzer, M. Mann, and K. Aktories, Nature 387:725-729, 1997; G. Flatau, E. Lemichez, M. Gauthier, P. Chardin, S. Paris, C. Fiorentini, and P. Boquet, Nature 387:729-733, 1997). Here we report that in addition to RhoA, Cdc42 and Rac also are targets for CNF1 in vitro and in intact cells. Treatment of HeLa cells with CNF1 induced a transient formation of microspikes and formation of membrane ruffles. CNF1 caused a transient 10- to 50-fold increase in the activity of the c-Jun N-terminal kinase. Tryptic peptides of Cdc42 obtained from CNF1-treated cells by immunoprecipitation exhibited an increase in mass of 1 Da compared to control peptides, indicating the deamidation of glutamine 61 by the toxin. The same increase in mass was observed with the respective peptides obtained from CNF1-modified recombinant Cdc42 and Rac1. Modification of recombinant Cdc42 and Rac1 by CNF1 inhibited intrinsic and GAP-stimulated GTPase activities and retarded binding of 2'(3')-O-(N-methylanthraniloyl)GDP. The data suggest that recombinant as well as cellular Cdc42 and Rac are substrates for CNF1.

MeSH Terms
Bacterial Toxins/genetics,metabolism,pharmacology Base Sequence Calcium-Calmodulin-Dependent Protein Kinases/metabolism Cell Cycle Proteins/genetics,metabolism Cytotoxins/genetics,metabolism,pharmacology DNA, Complementary Enzyme Activation Escherichia coli/metabolism Escherichia coli Proteins GTP-Binding Proteins/genetics,metabolism HeLa Cells Humans JNK Mitogen-Activated Protein Kinases Mass Spectrometry Mitogen-Activated Protein Kinases Molecular Sequence Data Recombinant Fusion Proteins/genetics,metabolism,pharmacology cdc42 GTP-Binding Protein rac GTP-Binding Proteins
Chemicals
Bacterial Toxins Cell Cycle Proteins Cytotoxins DNA, Complementary Escherichia coli Proteins Recombinant Fusion Proteins cytotoxic necrotizing factor type 1 Calcium-Calmodulin-Dependent Protein Kinases JNK Mitogen-Activated Protein Kinases Mitogen-Activated Protein Kinases GTP-Binding Proteins cdc42 GTP-Binding Protein rac GTP-Binding Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lerm M
Institut für Pharmakologie und Toxikologie der Albert-Ludwigs-Universität Freiburg, 79104 Freiburg, Germany.
Selzer J
Hoffmeyer A
Rapp U R
Aktories K
Schmidt G
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.67.2.496-503.1999
Published
1999-02-00
Pages
496-503
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC96347
Subset
IM
Databases
GENBANK
X70670
External Links
PubMed source DOI: 10.1128/IAI.67.2.496-503.1999 PMC Full Text View journal details
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