A C-terminal domain targets the Pseudomonas aeruginosa cytotoxin ExoU to the plasma membrane of host cells

被引:43
作者
Rabin, SDP
Veesenmeyer, JL
Bieging, KT
Hauser, AR
机构
[1] Northwestern Univ, Dept Microbiol Immunol, Chicago, IL 60611 USA
[2] Northwestern Univ, Dept Med, Chicago, IL 60611 USA
关键词
D O I
10.1128/IAI.74.5.2552-2561.2006
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
ExoU, a phospholipase injected into host cells by the type III secretion system of Pseudomonas aeruginosa, leads to rapid cytolytic cell death. Although the importance of ExoU in infection is well established, the mechanism by which this toxin kills host cells is less clear. To gain insight into how ExoU causes cell death, we examined its subcellular localization following transfection or type III secretion/translocation into HeLa cells. Although rapid cell lysis precluded visualization of wild-type ExoU by fluorescence microscopy, catalytically inactive toxin was readily detected at the periphery of HeLa cells. Biochemical analysis confirmed that ExoU was targeted to the membrane fraction of transfected cells. Visualization of ExoU peptides fused with green fluorescent protein indicated that the domain responsible for this targeting was in the C terminus of ExoU, between residues 550 and 687. Localization to the plasma membrane occurred within I h of expression, which is consistent with the kinetics of cytotoxicity. Together, these results indicate that a domain between residues 550 and 687 of ExoU targets this toxin to the plasma membrane, a process that may be important in cytotoxicity.
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页码:2552 / 2561
页数:10
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共 61 条
[11]  
Finck-Barbançon V, 1998, J BACTERIOL, V180, P6224
[12]   Multiple domains are required for the toxic activity of Pseudomonas aeruginosa ExoU [J].
Finck-Barbançon, V ;
Frank, DW .
JOURNAL OF BACTERIOLOGY, 2001, 183 (14) :4330-4344
[13]   ExoU expression by Pseudomonas aeruginosa correlates with acute cytotoxicity and epithelial injury [J].
FinckBarbancon, V ;
Goranson, J ;
Zhu, L ;
Sawa, T ;
WienerKronish, JP ;
Fleiszig, SMJ ;
Wu, C ;
MendeMueller, L ;
Frank, DW .
MOLECULAR MICROBIOLOGY, 1997, 25 (03) :547-557
[14]   Relationship between cytotoxicity and corneal epithelial cell invasion by clinical isolates of Pseudomonas aeruginosa [J].
Fleiszig, SMJ ;
Zaidi, TS ;
Preston, MJ ;
Grout, M ;
Evans, DJ ;
Pier, GB .
INFECTION AND IMMUNITY, 1996, 64 (06) :2288-2294
[15]   The exoenzyme S regulon of Pseudomonas aeruginosa [J].
Frank, DW .
MOLECULAR MICROBIOLOGY, 1997, 26 (04) :621-629
[16]   GENETIC AND PHYSICAL MAPPING OF THE PATATIN GENES IN POTATO AND TOMATO [J].
GANAL, MW ;
BONIERBALE, MW ;
ROEDER, MS ;
PARK, WD ;
TANKSLEY, SD .
MOLECULAR & GENERAL GENETICS, 1991, 225 (03) :501-509
[17]   The Yersinia YpkA Ser/Thr kinase is translocated and subsequently targeted to the inner surface of the HeLa cell plasma membrane [J].
Hakansson, S ;
Galyov, EE ;
Rosqvist, R ;
WolfWatz, H .
MOLECULAR MICROBIOLOGY, 1996, 20 (03) :593-603
[18]   Pseudomonas aeruginosa induces type-III-secretion-mediated apoptosis of macrophages and epithelial cells [J].
Hauser, AR ;
Engel, JN .
INFECTION AND IMMUNITY, 1999, 67 (10) :5530-5537
[19]   PepA, a secreted protein of Pseudomonas aeruginosa, is necessary for cytotoxicity and virulence [J].
Hauser, AR ;
Kang, PJ ;
Engel, JN .
MOLECULAR MICROBIOLOGY, 1998, 27 (04) :807-818
[20]   Type III protein secretion is associated with poor clinical outcomes in patients with ventilator-associated pneumonia caused by Pseudomonas aeruginosa [J].
Hauser, AR ;
Cobb, E ;
Bodí, M ;
Mariscal, D ;
Vallés, J ;
Engel, JN ;
Rello, J .
CRITICAL CARE MEDICINE, 2002, 30 (03) :521-528