Adenovirus E1b-58 kD antigen binds to p53 during infection of rodent cells: evidence for an N-terminal binding site on p53.
We show using mild extraction procedures that the p53 proto-oncogene forms a complex with adenovirus 5 E1b-58 kD during infection. These complexes are detected as coimmunoprecipitates from radiolabeled extracts of adenovirus infected cells on SDS-PAGE. Furthermore, adenovirus mutants with defects in E1b-58 kD fail to form complexes, whereas mutants in other early region genes still show evidence of complex. Using a panel of monoclonal antibodies to mouse p53, we show that antibodies reacting with N-terminal epitopes on p53, displace E1b-58 kD. This result suggests that E1b-58 kD binds to an N-terminal region of mouse p53. In addition, in a transient transfection assay in monkey COS cells, we show that an N-terminal deletion mutant of mouse p53 does not bind to E1b-58 kD but wild-type mouse p53 does bind. This result again suggests that E1b-58 kD binds an N-terminal determinant on p53.
Braithwaite
A W
AW
Division of Cell Biology, John Curtin School of Medical Research, Australian National University, Canberra.
Blair
G E
GE
Nelson
C C
CC
McGovern
J
J
Bellett
A J
AJ
D000256:Adenoviridae / Q000235:genetics*; D017456:Adenovirus Early Proteins; D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002460:Cell Line; D002872:Chromosome Deletion; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D008970:Molecular Weight; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D000090063:Proto-Oncogene Mas; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D017456:Adenovirus Early Proteins; D000911:Antibodies, Monoclonal; C000715470:MAS1 protein, human; D009856:Oncogene Proteins, Viral; D000090063:Proto-Oncogene Mas; D016159:Tumor Suppressor Protein p53