Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A.
We have purified the 36 and 63 kd cellular proteins known to associate with polyomavirus middle and small tumor (T) antigens and SV40 small t antigen. Microsequencing of the 36 kd protein indicated that it was probably identical to the catalytic subunit of protein phosphatase 2A (PP2A). Identity was confirmed by comigration on two-dimensional (2D) gels and by 2D analysis of complete chymotryptic digests. In addition, PP2A-like phosphatase activity was detected in immunoprecipitates of wild-type middle T. Immunoblotting experiments, comigration on 2D gels, and 2D analysis of limit chymotryptic digests demonstrated that the 63 kd protein, present in the middle T complex in approximately equimolar ratio to the 36 kd protein, is a known regulatory subunit of the PP2A holoenzyme. Finally, the 36 kd PP2A catalytic subunit can be immunoprecipitated by anti-pp60c-src antisera only from cells expressing wild-type middle T. These results suggest that complex formation between PP2A and T antigens may be important for T antigen-mediated transformation.
Pallas
D C
DC
Division of Cellular and Molecular Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115.
Shahrik
L K
LK
Martin
B L
BL
Jaspers
S
S
Miller
T B
TB
Brautigan
D L
DL
Roberts
T M
TM
D000595:Amino Acid Sequence; D000818:Animals; D000952:Antigens, Polyomavirus Transforming; D001665:Binding Sites; D002460:Cell Line; D015151:Immunoblotting; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism*; D010765:Phosphorylase Phosphatase / Q000378:metabolism; D011120:Polyomavirus / Q000276:immunology*; D011485:Protein Binding; D054648:Protein Phosphatase 2; D013539:Simian virus 40 / Q000276:immunology*; D014357:Trypsin
D000952:Antigens, Polyomavirus Transforming; D046911:Macromolecular Substances; D010446:Peptide Fragments; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D010765:Phosphorylase Phosphatase; D014357:Trypsin