Mechanism underlying inhibition of intestinal apical Cl-/OH- exchange following infection with enteropathogenic E-coli

被引:102
作者
Gill, Ravinder K.
Borthakur, Alip
Hodges, Kim
Turner, Jerrold R.
Clayburgh, Daniel R.
Saksena, Seema
Zaheer, Ayesha
Ramaswamy, Krishnamurthy
Hecht, Gail
Dudeja, Pradeep K.
机构
[1] Univ Illinois, Med Res Ctr, Jesse Brown VA Med Ctr, Chicago, IL 60612 USA
[2] Univ Illinois, Sect Digest Dis & Nutr, Dept Med, Chicago, IL USA
[3] Univ Chicago, Dept Pathol, Chicago, IL 60637 USA
关键词
D O I
10.1172/JCI29625
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Enteropathogenic E. coli (EPEC) is a major cause of infantile diarrhea, but the pathophysiology underlying associated diarrhea is poorly understood. We examined the role of the luminal membrane Cl-/OH- exchange process in EPEC pathogenesis using in vitro and in vivo models. Cl-/OH- exchange activity was measured as OH- gradient-driven Cl-36(-) uptake. EPEC infection (60 minutes-3 hours) inhibited apical Cl-/OH- exchange activity in human intestinal Caco-2 and T84 cells. This effect was dependent upon the bacterial type III secretory system (TTSS) and involved secreted effector molecules EspG and EspG2, known to disrupt the host microtubular network. The microtubule-disrapting agent colchicine (100 mu M, 3 hours) also inhibited Cl-36- uptake. The plasma membrane expression of major apical anion exchanger DRA (SLC26A3) was considerably reduced in EPEC-infected cells, corresponding with decreased Cl-/OH- exchange activity. Confocal microscopic studies showed that EPEC infection caused a marked redistribution of DRA from the apical membrane to intraceflular compartments. Interestingly, infection of cells with an EPEC mutant deficient in espG significantly attenuated the decrease in surface expression of DRA protein as compared with treatment with wild-type EPEC. EPEC infection in vivo (1 day) also caused marked redistribution of surface DRA protein in the mouse colon. Our data demonstrate that EspG and EspG2 play an important role in contributing to EPEC infection-associated inhibition of luminal membrane chloride transport via modulation of surface DRA expression.
引用
收藏
页码:428 / 437
页数:10
相关论文
共 47 条
[1]   C-terminal domains of Na+/H+ exchanger isoform 3 are involved in the basal and serum-stimulated membrane trafficking of the exchanger [J].
Akhter, S ;
Cavet, ME ;
Tse, CM ;
Donowitz, M .
BIOCHEMISTRY, 2000, 39 (08) :1990-2000
[2]   Enteropathogenic Escherichia coli inhibits butyrate uptake in Caco-2 cells by altering the apical membrane MCT1 level [J].
Borthakur, A ;
Gill, RK ;
Hodges, K ;
Ramaswamy, K ;
Hecht, G ;
Dudeja, PK .
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY, 2006, 290 (01) :G30-G35
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   Human DRA functions as a sulfate transporter in Sf9 insect cells [J].
Byeon, MK ;
Frankel, A ;
Papas, TS ;
Henderson, KW ;
Schweinfest, CW .
PROTEIN EXPRESSION AND PURIFICATION, 1998, 12 (01) :67-74
[5]   Enteropathogenic Escherichia coli (EPEC) attachment to epithelial cells:: exploiting the host cell cytoskeleton from the outside [J].
Celli, J ;
Deng, WY ;
Finlay, BB .
CELLULAR MICROBIOLOGY, 2000, 2 (01) :1-9
[6]   Acute regulation of the SLC26A3 congenital chloride diarrhoea anion exchanger (DRA) expressed in Xenopus oocytes [J].
Chernova, MN ;
Jiang, LW ;
Shmukler, BE ;
Schweinfest, CW ;
Blanco, P ;
Freedman, SD ;
Stewart, AK ;
Alper, SL .
JOURNAL OF PHYSIOLOGY-LONDON, 2003, 549 (01) :3-19
[7]   N-glycosylation and microtubule integrity are involved in apical targeting of prostate-specific membrane antigen:: implications for immunotherapy [J].
Christiansen, JJ ;
Rajasekaran, SA ;
Inge, L ;
Cheng, LR ;
Anilkumar, G ;
Bander, NH ;
Rajasekaran, AK .
MOLECULAR CANCER THERAPEUTICS, 2005, 4 (05) :704-714
[8]   Rapid modulation of electrolyte transport in Caco-2 cell monolayers by enteropathogenic Escherichia coli (EPEC) infection [J].
Collington, GK ;
Booth, IW ;
Knutton, S .
GUT, 1998, 42 (02) :200-207
[9]   The filamentous type III secretion translocon of enteropathogenic Escherichia coli [J].
Daniell, SJ ;
Takahashi, N ;
Wilson, R ;
Friedberg, D ;
Rosenshine, I ;
Booy, FP ;
Shaw, RK ;
Knutton, S ;
Frankel, G ;
Aizawa, S .
CELLULAR MICROBIOLOGY, 2001, 3 (12) :865-871
[10]   A 2ND CHROMOSOMAL GENE NECESSARY FOR INTIMATE ATTACHMENT OF ENTEROPATHOGENIC ESCHERICHIA-COLI TO EPITHELIAL-CELLS [J].
DONNENBERG, MS ;
YU, J ;
KAPER, JB .
JOURNAL OF BACTERIOLOGY, 1993, 175 (15) :4670-4680