Involvement of ADP-ribosylation factor 1 in cholera toxin induced morphological changes of Chinese hamster ovary cells

被引:17
作者
Morinaga, N
Kaihou, Y
Vitale, N
Moss, J
Noda, M
机构
[1] Chiba Univ, Sch Med, Dept Microbiol 2, Chuo Ku, Chiba 2608670, Japan
[2] INSERM U338, Ctr Neurochim, F-67084 Strasbourg, France
[3] NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA
关键词
D O I
10.1074/jbc.M101184200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
ADP-ribosylation factor 1 (ARF1) was originally found as a cofactor in CT-catalyzed ADP-ribosylation of Ga, but is now known to participate in vesicle trafficking. We asked whether ARF1 function in vesicular trafficking is necessary for CT induced morphological changes in Chinese hamster ovary (CHO) cells, which result from increased intracellular cAMCP. Brefeldin A treatment of cells suppressed CT action, confirming a requirement for Golgi integrity. Overexpression of a GFP-ARF1 fusion protein did not affect the morphological changes induced by CT, but changes were reduced in cells over expressing guanine nucleotide exchange-defective ARF1(T31N) or GTP hydrolysis-deficient ARF1(Q71L) mutants. In cells expressing these mutants, 8-bromo-cAMP induced changes similar to those seen in cells transfected with ARF1 or vector. Inhibition of CT action was specific for mutants of ARF1 and not reproduced by analogous mutants of ARF5 or ARF6. ARF1(Q71L) was mostly colocalized with beta COP, but ARF5(Q71L) less so. ARF6(Q67L) did not colocalize with beta COP and was partially associated with the plasma membrane. These data are consistent with the conclusion that ARF1 influenced CT action in cells by its specific function in the vesicular transport pathway used by CT to travel from plasma membrane to Golgi to ER.
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页码:22838 / 22843
页数:6
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