RAF-1 PROTEIN-KINASE IS REQUIRED FOR GROWTH OF INDUCED NIH/3T3 CELLS
RAF-1 PROTEIN-KINASE IS REQUIRED FOR GROWTH OF INDUCED NIH/3T3 CELLS
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DOI：
10.1038/349426a0
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发表时间：
1991-01-31
期刊：
影响因子：
64.8
通讯作者：
RAPP, UR
中科院分区：
文献类型：
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作者：
KOLCH, W;HEIDECKER, G;RAPP, UR
许多生长因子调节细胞质RAF-1蛋白激酶1-10，这与其在生长信号转导中具有中心作用一致。 该激酶广泛表达11，并可促进增殖12，可能以依赖于生长因子受体和膜相关癌基因的方式13-15。 我们现在检查了血清和TPa（12-O-十四烷酰基佛波醇-13-乙酸酯）调节的NIH/3 T3细胞生长对RAF-1激酶的依赖性，以确定Raf-1是否对受体信号传导至关重要。 我们通过表达c-raf-1反义RNA或激酶缺陷的c-raf-1突变体来抑制Raf-1功能。 c-raf-1反义RNA干扰正常NIH/3 T3细胞增殖并逆转raf转化细胞 在回复突变体细胞中，血清或TPA诱导的DNA复制被消除或减少成比例的Raf蛋白水平的降低。 激酶缺陷的Raf-1突变体（RAF 301）或调节结构域片段（HCR）的表达抑制血清诱导的NIH/3 T3细胞增殖和raf转化甚至更有效。反义RNA或反义RNA 301抑制Ki-和Ha-ras癌基因的增殖和转化。 我们的结论是，raf功能作为一个重要的信号转导下游的血清生长因子受体，蛋白激酶C和ras。
MANY growth factors regulate the cytoplasmic RAF-1 protein kinase 1-10, consistent with its having a central role in transduction of growth signals. The kinase is ubiquitously expressed 11 and can promote proliferation 12, presumably in a manner dependent on growth-factor receptors and membrane-associated oncogenes 13-15. We have now examined the dependence of serum- and TPa (12-O-tetradecanoylphorbol-13-acetate)-regulated NIH/3T3 cell growth on RAF-1 kinase to determine whether Raf-1 is essential for receptor signalling. We inhibited Raf-1 function by expressing c-raf-1 antisense RNA or kinase-defective c-raf-1 mutants. Antisense RNA for c-raf-1 interferes with proliferation of normal NIH/3T3 cells and reverts raf-transformed cells. In revertant cells, DNA replication induced by serum or TPA was eliminated or reduced proportionately to the reduction in Raf protein levels. Expression of a kinase-defective Raf-1 mutant (craf301) or a regulatory domain fragment (HCR) inhibited serum-induced NIH/3T3-cell proliferation and raf transformation even more efficiently. Inhibition by antisense RNA or craf301 blocked proliferation and transformation by Ki- and Ha-ras oncogenes. We conclude that raf functions as an essential signal transducer downstream of serum growth factor receptors, protein kinase C and ras.