CHARACTERIZATION OF THE INTERACTION OF POLYOMAVIRUS MIDDLE T-ANTIGEN WITH TYPE-2A PROTEIN PHOSPHATASE

被引:38
作者
ULUG, ET
CARTWRIGHT, AJ
COURTNEIDGE, SA
机构
[1] EUROPEAN MOLEC BIOL LAB,DIFFERENTIAT PROGRAMME,MEYERHOFSTR 1,W-6900 HEIDELBERG,GERMANY
[2] KANSAS STATE UNIV AGR & APPL SCI,DIV BIOL,VIROL & ONCOL SECT,MANHATTAN,KS 66506
关键词
D O I
10.1128/JVI.66.3.1458-1467.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Two cellular proteins of 36 and 63 kDa which bind the small T and middle T antigens of polyomavirus recently have been identified as the catalytic and regulatory subunits of the phosphoserine/threonine-specific type 2A protein phosphatase (PP2A). We report here the presence of phosphoseryl protein phosphatase activity associated with polyomavirus small T and middle T antigens in immunoprecipitates prepared from virus-infected and transformed cells. Phosphatase activity was also found associated with middle T-antigen mutants, some of which had been defined previously to associate with 36- and 63-kDa cellular proteins. Middle T-antigen-associated phosphatase activity was sensitive to okadaic acid and microcystin-LR, inhibitors of PP2A, and insensitive to inhibitor 1 or 2, orthovanadate, or EDTA. Using antiserum specific for the catalytic subunit of PP2A, we found that unlike the majority of PP2A, middle T-antigen-bound PP2A was membrane associated. However, no gross change in the amount, activity, or localization of PP2A could be attributed to middle T-antigen expression in transformed cells. Anti-PP2A antibodies coprecipitated a 63-kDa protein from normal cells and in addition coprecipitated middle T antigen, 60- and 61-kDa proteins (identified as src family members), and an 81-kDa protein from middle T-antigen-transformed cells. Furthermore, we detected protein kinase activity in PP2A immunoprecipitates and protein phosphatase activity in src immune complexes from extracts of middle T-antigen-transformed, but not normal, cells. These results reinforce the notion that at least a portion of middle T antigen bridges a protein kinase with a protein phosphatase.
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页码:1458 / 1467
页数:10
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