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J. Biol. Chem., Vol. 265, Issue 12, 6569-6575, Apr, 1990

Direct stimulation of adenylate cyclase by mechanical forces in S49 mouse lymphoma cells during hyposmotic swelling

PA Watson
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822.

S49 mouse lymphoma cells respond to swelling deformation with rapid increases in intracellular calcium and cAMP. Experiments demonstrate that these increases in calcium and cAMP concentrations are not coupled in a regulatory manner. Direct inhibition of adenylate cyclase in wild type cells with miconazole prevented swelling-induced accumulation of cAMP. No effect of swelling was observed on the activity of cAMP phosphodiesterase. Additionally, complete inhibition of cAMP phosphodiesterase did not prevent swelling-induced cAMP accumulation. Experiments involving cyc- mutants (lacking the Gs-alpha protein) and 2',5'-dideoxyadenosine indicate that increased adenylate cyclase activity with swelling is not mediated by Gs. No evidence was found for attenuation of Gi-mediated inhibition of adenylate cyclase activity following swelling. In addition, exposure to pertussis toxin or phorbol ester, which disrupts Gi inhibition of adenylate cyclase did not prevent cAMP accumulation following swelling. Disruption of the actin membrane skeleton resulted in a significant accumulation of cAMP which was not further increased by swelling. Disruption of the microtubular cytoskeleton also increased cAMP content in S49 cells which could be further increased by swelling. It is concluded that S49 cell-adenylate cyclase responds directly to mechanical forces transmitted through the actin membrane skeleton.
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