TRANSFORMATION-DEFECTIVE MUTANTS OF POLYOMAVIRUS MIDDLE T-ANTIGEN ASSOCIATE WITH PHOSPHATIDYLINOSITOL 3-KINASE (PI 3-KINASE) BUT ARE UNABLE TO MAINTAIN WILD-TYPE LEVELS OF PI 3-KINASE PRODUCTS IN INTACT-CELLS

被引:63
作者
LING, LE
DRUKER, BJ
CANTLEY, LC
ROBERTS, TM
机构
[1] HARVARD UNIV,SCH MED,DANA FARBER CANC INST,BOSTON,MA 02115
[2] TUFTS UNIV,SCH MED,DEPT PHYSIOL,BOSTON,MA 02111
关键词
D O I
10.1128/JVI.66.3.1702-1708.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Middle T antigen (MT) of polyomavirus causes transformation by associating with a number of cellular proteins. The association with and activation of two such proteins, phosphatidylinositol 3-kinase (PI 3-kinase) and pp60c-src, appears to be necessary for transformation by MT. The tyrosine kinase activity of MT-associated pp60c-src is significantly increased when assayed in vitro, and levels of phosphotyrosine-containing proteins are elevated in vivo. Similarly, levels of the PI 3-kinase products phosphatidylinositol-3,4-bisphosphate [PI(3,4)P2] and phosphatiylinositol-3,4,5-trisphosphate [PI(3,4,5)P3] are constitutively elevated in MT-transformed cells. However, the formation of a complete MT/cellular protein complex and the activation of tyrosine kinase are not sufficient to cause transformation, since the transformation-defective mutants 248m and dl1015 associate with all wild-type MT-associated proteins, including PI 3-kinase and pp60c-src, and neither mutant appears to be defective in MT-associated tyrosine kinase activity. Studies presented here compared (i) the amount of PI 3-kinase activity associated with the MT complex and (ii) levels of [H-3]inositol incorporation into PI 3-kinase products in cells expressing mutant or wild-type MT. The results show that dl1015 is defective in both assays, whereas 248m is defective only for incorporation of [H-3]inositol into PI(3,4,5)P2 and PI(3,4)P3. These findings identify a biochemical defect in the 248m mutant and corroborate previous results correlating transformation and elevated levels of PI 3-kinase products in vivo. In addition, they indicate that PI 3-kinase product levels are affected by factors other than simply the amount of PI 3-kinase activity associated with the MT complex.
引用
收藏
页码:1702 / 1708
页数:7
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