Cell fractionation in non-ionic detergent distinguishes sub-populations of polyoma virus middle T antigen and reveals a novel form.
The transforming function of polyoma virus, middle T antigen (MT), interacts with several cellular enzymes, essential to its oncogenic activity. We have used cell fractionation to study the various MT/cellular protein complexes. We demonstrate that MT can be separated into three sub-species, dependent upon extraction in two buffers that we designate A and B: Antigen extracted from whole cells by both buffers (called MT1) is associated with most of the phosphorylated phosphatidyl-inositol kinase 85 kD subunit, pp85, and protein phosphatase 2A. Antigen (MT2), associated with the greater portion of pp60c-src, is extracted by buffer B, but not buffer A. A third population (MT3), resistant to extraction by either buffer, is not detectably associated with protein phosphatase 2A or pp85. It is, however, associated with a low level kinase activity. The interaction between pp60c-src and MT appears to influence the formation of both MT2 and MT3. MT2 fractionates with the cellular microtubule network, but does not appear to be directly associated with it. MT3, a previously undescribed population, comprises about one third of MT in wild type antigen-containing cells. It is missing in mutants incapable of interacting with pp60c-src, but exists in the absence of an interaction with pp85. We suggest that MT3 may be an intermediate in, or product of, one of the MT/pp60c-src signalling pathways, distinct from that involving pp85.
Krauzewicz
N
N
Department of Virology, Royal Postgraduate Medical School, London, UK.
Elliott
J
J
Griffin
B E
BE
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000302:isolation & purification*; D002458:Cell Fractionation; D003902:Detergents / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000302:isolation & purification; D011120:Polyomavirus / Q000276:immunology*; D054648:Protein Phosphatase 2; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000302:isolation & purification; D051381:Rats; D014404:Tubulin / Q000302:isolation & purification
D000952:Antigens, Polyomavirus Transforming; D003902:Detergents; D014404:Tubulin; D017853:Phosphotransferases (Alcohol Group Acceptor); C020077:1-phosphatidylinositol-4-phosphate 5-kinase; D016392:Proto-Oncogene Proteins pp60(c-src); D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2