A dominant negative RAS-specific guanine nucleotide exchange factor reverses neoplastic phenotype in K-ras transformed mouse fibroblasts

被引:0
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作者
Paola Bossù
Marco Vanoni
Valeria Wanke
Maria Paola Cesaroni
Franco Tropea
Gabriella Melillo
Cinzia Asti
Stefano Porzio
Paolo Ruggiero
Vito Di Cioccio
Giovanni Maurizi
Annibale Ciabini
Lilia Alberghina
机构
[1] Università degli Studi di Milano-Bicocca,Dipartimento di Biotecnologie e Bioscienze
[2] Centro Ricerche Dompé,undefined
来源
Oncogene | 2000年 / 19卷
关键词
CDC25; signal transduction therapy; -luciferase; oncogenes; GEF;
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学科分类号
摘要
Ras proteins are small GTPases playing a pivotal role in cell proliferation and differentiation. Their activation state depends on the competing action of GTPase Activating Proteins (GAP) and Guanine nucleotide Exchange Factors (GEF). A tryptophan residue (Trp1056 in CDC25Mm-GEF), conserved in all ras-specific GEFs identified so far has been previously shown to be essential for GEF activity. Its substitution with glutamic acid results in a catalytically inactive mutant, which is able to efficiently displace wild-type GEF from p21ras and to originate a stable ras/GEF binary complex due to the reduced affinity of the nucleotide-free ras/GEF complex for the incoming nucleotide. We show here that this ‘ras-sequestering property' can be utilized to attenuate ras signal transduction pathways in mouse fibroblasts transformed by oncogenic ras. In fact over-expression of the dominant negative GEFW1056E in stable transfected cells strongly reduces intracellular rasGTP levels in k-ras transformed fibroblasts. Accordingly, the transfected fibroblasts revert to wild-type phenotype on the basis of morphology, cell cycle and anchorage independent growth. The reversion of the transformed phenotype is accompanied by DNA endoreduplication. The possible use of dominant negative ras-specific GEFs as a tool to down-regulate tumor growth is discussed.
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页码:2147 / 2154
页数:7
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