The membrane localization domain is required for intracellular localization and autoregulation of YopE in Yersinia pseudotuberculosis.
Recent work has shown that a domain of YopE of Yersinia pseudotuberculosis ranging from amino acids 54 to 75 (R. Krall, Y. Zhang, and J. T. Barbieri, J. Biol. Chem. 279:2747-2753, 2004) is required for proper localization of YopE after ectopic expression in eukaryotic cells. This domain, called the membrane localization domain (MLD), has not been extensively studied in Yersinia. Therefore, an in cis MLD deletion mutant of YopE was created in Y. pseudotuberculosis. The mutant was found to secrete and translocate YopE at wild-type levels. However, the mutant was defective in the autoregulation of YopE expression after the infection of HeLa cells. Although the mutant translocated YopE at wild-type levels, it showed a delayed HeLa cell cytotoxicity. This delay was not caused by a change in GTPase activating protein (GAP) activity, since the mutant showed wild-type YopE GAP activity toward Rac1 and RhoA. The MLD mutant displayed a changed intracellular localization pattern of YopE in HeLa cells after infection, and the YopEDeltaMLD protein was found to be dispersed within the whole cell, including the nucleus. In contrast, wild-type YopE was found to localize to the perinuclear region of the cell and was not found in the nucleus. In addition, the yopEDeltaMLD mutant was avirulent. Our results suggest that YopE must target proteins other than RhoA and Rac1 and that the MLD is required for the proper targeting and hence virulence of YopE during infection. Our results raise the question whether YopE is a regulatory protein or a "true" virulence effector protein.
Isaksson
Elin L
EL
Department of Molecular Biology, Laboratory of Molecular Infectious Medicine, Umeå University, S-90187 Umeå, Sweden.
Aili
Margareta
M
Fahlgren
Anna
A
Carlsson
Sara E
SE
Rosqvist
Roland
R
Wolf-Watz
Hans
H
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D021381:Protein Transport / Q000502:physiology; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D015012:Yersinia pseudotuberculosis Infections / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; C063239:yopE protein, Yersinia
Infect Immun. 2001 May;69(5):3516-8
11292787
Infect Immun. 2005 Dec;73(12):7938-45
16299285
Int J Med Microbiol. 2000 Oct;290(4-5):381-7
11111915
Curr Top Microbiol Immunol. 2005;291:147-66
15984080
Methods Enzymol. 2002;358:359-70
12474400
J Biol Chem. 2004 Jan 23;279(4):2747-53
14597627
Mol Microbiol. 1995 May;16(4):635-47
7476159
Cell Microbiol. 2006 Jun;8(6):1020-33
16681842
J Biol Chem. 2007 Apr 27;282(17):13022-32
17311921
Mol Microbiol. 2000 May;36(3):737-48
10844661
Cell Microbiol. 2001 May;3(5):301-10
11298653
Microb Pathog. 2003 Jun;34(6):297-308
12782482
Int J Med Microbiol. 2008 Apr;298(3-4):183-92
17597003
Infect Immun. 1982 Aug;37(2):506-12
6749681
J Bacteriol. 1996 Dec;178(24):7227-33
8955406
Infect Immun. 1995 Jun;63(6):2269-76
7768608
Mol Microbiol. 2000 Aug;37(3):515-27
10931345
Mol Microbiol. 1998 Aug;29(3):799-813
9723919
Infect Immun. 1991 Dec;59(12):4562-9
1937815
Traffic. 2008 Nov;9(11):1948-57
18778330
Mol Microbiol. 2002 Dec;46(5):1381-90
12453223
Biochem Biophys Res Commun. 2002 Feb 15;291(1):91-100
11829467
J Bacteriol. 2008 Apr;190(8):2726-38
18039770
EMBO J. 1994 Feb 15;13(4):964-72
8112310
Mol Microbiol. 1990 Apr;4(4):657-67
2191183
J Mol Biol. 1983 Jun 5;166(4):557-80
6345791
Mol Cell. 2002 May;9(5):971-80
12049734
Nat Rev Microbiol. 2005 May;3(5):397-410
15821726
J Bacteriol. 1996 Mar;178(5):1310-9
8631707
Mol Microbiol. 1997 Apr;24(1):73-91
9140967
Eur J Cell Biol. 2000 Oct;79(10):659-71
11089914
EMBO J. 2001 Oct 1;20(19):5373-82
11574469
PLoS Pathog. 2008 Jan;4(1):e3
18193942
Annu Rev Microbiol. 2005;59:69-89
15847602
Infect Immun. 1984 Jan;43(1):72-8
6317574
EMBO J. 1999 Feb 1;18(3):578-85
9927417
J Cell Biol. 1999 Nov 29;147(5):1009-22
10579721