Expression and complex formation of simian virus 40 large T antigen and mouse p53 in insect cells.
Recombinant baculoviruses were constructed which express simian virus 40 large T antigen (SVT-Ag) or murine p53 to high levels in infected insect cells. Characterization of the expressed proteins revealed that they display many properties of the corresponding mammalian-derived proteins. Both proteins are of wild-type size, localize to the nucleus, are recognized by several SVT-Ag- or p53-specific monoclonal antibodies, and are phosphorylated in this system. Complexes are formed between baculovirus-derived SVT-Ag and p53 after coinfection of insect cells with both recombinant viruses. After infection of insect cells with either virus individually, each protein can self-associate to form a variety of oligomeric species. Pulse-chase experiments indicated that both SVT-Ag and p53 are highly stable in insect cells, even in the absence of complex formation.
O'Reilly
D R
DR
Department of Entomology, University of Georgia, Athens 30602.
Miller
L K
LK
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D007118:Immunoassay; D007304:Insect Viruses / Q000235:genetics; D007700:Kinetics; D051379:Mice; D009036:Moths; D009363:Neoplasm Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics / Q000276:immunology*; D016159:Tumor Suppressor Protein p53
D000952:Antigens, Polyomavirus Transforming; D000939:Epitopes; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D016159:Tumor Suppressor Protein p53