Protein synthesis is required for cholera toxin-induced stimulation of arachidonic acid metabolism.
The molecular events in the mechanism of action of cholera toxin were analyzed using Chinese hamster ovary (CHO) cells. Cholera toxin stimulated both 3',5'-cyclic adenosine monophosphate (cAMP) synthesis and arachidonic acid metabolism in these cells. The turnover of phospholipid by cholera toxin-induced stimulation of phospholipase activity evoked the synthesis of PGE2 and other prostaglandins. Cholera toxin-induced release of both [3H]arachidonic acid and PGE2 was blocked by addition of either cycloheximide or actinomycin D. In contrast, accumulation of cAMP in cholera toxin-treated CHO cells was unaffected by adding these drugs. Further, dibutyryl cAMP or forskolin caused [3H]arachidonic acid release, which also was blocked by cycloheximide and actinomycin D. We concluded that the sequence of molecular events in cholera toxin-treated CHO cells first involved activation of adenylate cyclase, which caused an increase in cAMP. In turn, cAMP promoted transcription of mRNA that encoded either a specific phospholipase or a phospholipase-activating protein. The emerging arachidonic acid metabolites (e.g., PGE2 and PGF2 alpha) might be important mediators of cholera toxin's stimulatory effects on vascular permeability and smooth muscle contraction in the intestine during cholera.
Peterson
J W
JW
Department of Microbiology, University of Texas Medical Branch, Galveston 77550.
Reitmeyer
J C
JC
Jackson
C A
CA
Ansari
G A
GA
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D002460:Cell Line / Q000166:cytology / Q000187:drug effects; D002772:Cholera Toxin / Q000494:pharmacology*; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000096:biosynthesis / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015232:Dinoprostone / Q000096:biosynthesis*; D014176:Protein Biosynthesis
D001095:Arachidonic Acids; D003609:Dactinomycin; D016718:Arachidonic Acid; D002772:Cholera Toxin; D003513:Cycloheximide; D000242:Cyclic AMP; D015232:Dinoprostone