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Am J Physiol Cell Physiol 268: C1460-C1466, 1995;
0363-6143/95 $5.00
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AJP - Cell Physiology, Vol 268, Issue 6 C1460-C1466, Copyright © 1995 by American Physiological Society

ARTICLES

Constitutively active mutant GS alpha (G225T) and null-mutant G alpha i-2 (G203T) induce primitive endoderm from stem cells

P. Gao, D. C. Watkins and C. C. Malbon
Department of Molecular Pharmacology, State University of New York-Stony Brook 11794-8651, USA.

In F9 teratocarcinoma stem cells, retinoic acid induces a primitive endoderm-like phenotype and a sharp decline in G alpha i-2, a response mimicked by expression of RNA antisense to G alpha i-2 in the absence of this morphogen (D. C. Watkins, G. L. Johnson, and C. C. Malbon. Science Wash. DC 258: 1373-1375, 1992). The role of the GS alpha/G alpha i-2 axis in cellular differentiation was explored. In the absence of retinoic acid, F9 stem cells stably expressing a constitutively active mutant of GS alpha (G225T) progressed to the primitive endoderm phenotype, as judged by morphological and differentiation markers, such as tissue plasminogen activator. Although elevated in cells expressing G225T GS alpha, adenosine 3',5'-cyclic monophosphate does not mimic retinoic acid action and alone fails to induce stem cells to primitive endoderm. In the absence of retinoic acid, expression of a null mutant of G alpha i-2 (G203T) also induced stem cells to primitive endoderm. These observations establish G proteins in the GS alpha/G alpha i-2 axis as a control point for regulating progression to primitive endoderm independent of adenylate cyclase, in the present study's model of early mouse development.


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