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ras ‐p21 Activates phospholipase D and A2, but not phospholipase C or PKC, in Xenopus laevis Oocytes
Author(s) -
Carnero Amancio,
Dolfi Fabrizio,
Lacal Juan Carlos
Publication year - 1994
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240540415
Subject(s) - xenopus , microinjection , phospholipase d , protein kinase c , microbiology and biotechnology , phospholipase c , biology , mapk/erk pathway , signal transduction , anti apoptotic ras signalling cascade , dna synthesis , staurosporine , biochemistry , in vitro , gene
Xenopus laevis oocytes are a powerful tool for the characterization of signal transduction pathways leading to the induction of DNA synthesis. Since activation of PLA2, PLC, or PLD has been postulated as a mediator of ras function, we have used the oocyte system to study the putative functional relationship between ras ‐p21 and these phospholipases. A rapid generation of PA and DAG was observed after ras ‐p21 microinjection, suggesting the activation of both PLC and PLD enzymes. However, production of DAG was sensitive to inhibition of the PA‐hydrolase by propranolol, indicating that PLD is the enzyme responsible for the generation of both PA and DAG. Microinjection of PLD or ras ‐p21 induced the late production of lysophosphatidylcholine on a p42 MAPK ‐dependent manner, an indication of the activation of a PLA2. Inhibition of this enzyme by quinacrine does not inhibit PLD‐ or ras ‐induced GVBD, suggesting that PLA2 activation is not needed for ras or PLD function. Contrary to 3T3 fibroblasts, where ras ‐p21 is functionally dependent for its mitogenic activity on TPA‐ and staurosporine‐sensitive PKC isoforms, in Xenopus oocytes, induction of GVBD by ras ‐p21 was independent of PKC, while PLC‐induced GVBD was sensitive to PKC inhibition. Thus, our results demonstrate the activation of PLD and PLA2 by ras ‐p21 proteins, while no effect on PLC was observed.