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Rapid stimulation of diacylglycerol production in Xenopus oocytes by microinjection of H-ras p21.

The p21 products of ras proto-oncogenes are thought to be important components in pathways regulating normal cell proliferation and differentiation. These proteins acquire transforming properties as a result of activating lesions that convert ras genes to oncogenes in a wide array of malignancies. In Xenopus laevis oocytes, microinjection of transforming ras p21 is a potent inducer of maturation, whereas microinjection of a monoclonal antibody to ras p21 inhibits normal maturation induced by hormones. The phosphoinositide pathway is a ubiquitous system that appears to play a key role in diverse cellular functions. By use of the Xenopus oocyte system, it was possible to quantitate the effects of ras p21 microinjection on individual components of the phosphoinositide pathway. Within 20 minutes of microinjection, levels of phosphatidylinositol 4,5-bisphosphate, inositol 1-phosphate, and inositol bisphosphate increased 1.5- to 2-fold. The most striking effects were on diacylglycerol, which increased 5-fold under the same conditions. In contrast, the normal ras p21 protein induced no detectable alteration in any of the metabolites analyzed. The earliest effects of the transforming p21 on phosphoinositol turnover were observable within 2 minutes, implying a very rapid effect of ras p21 on the enzymes involved in phospholipid metabolism.