A DNA replication-positive mutant of simian virus 40 that is defective for transformation and the production of infectious virions.
Simian virus 40 (SV40) mutant 5002 carries base pair substitutions of C-5109----T and C-5082----T. These mutations lie in a region of the genome that encodes amino acids common to the large and small viral tumor antigens (T and t antigens, respectively) and result in amino acid substitutions of Leu-19----Phe and Pro-28----Ser. In contrast to wild-type SV40, which produces large plaques that are clearly visible 8 days postinfection, mutant 5002 is defective for productive infection, producing tiny plaques that arise at around 21 days postinfection. However, 5002 is capable of replicating viral DNA and producing normal amounts of capsid proteins, indicating that the mutations alter an activity of T antigen that is required subsequent to DNA synthesis, such as maturation, viral assembly, or release of virions. The mutant T antigen has normal ATPase activity, is phosphorylated in a manner that is indistinguishable from that of the wild-type T antigen, and retains the ability to oligomerize. 5002 complements mutants defective in T antigen host range-adenovirus helper function for productive infection. Thus, T antigen encodes two activities that affect at least two different steps in viral infection other than DNA replication, one inactivated by mutations in the host range-adenovirus helper domain and one inactivated by the mutations present in 5002. The 5002-encoded T antigen is also defective for transformation of REF52 cells when expressed from the normal SV40 early promoter, although this defect can be partially overcome by expressing the protein from stronger promoters.
Peden
K W
KW
Howard Hughes Institute Laboratory, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Spence
S L
SL
Tack
L C
LC
Cartwright
C A
CA
Srinivasan
A
A
Pipas
J M
JM
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D003673:Defective Viruses / Q000276:immunology / Q000378:metabolism*; D005814:Genes, Viral; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010748:Phosphopeptides / Q000302:isolation & purification; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D010948:Viral Plaque Assay; D014771:Virion / Q000235:genetics* / Q000276:immunology
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D010748:Phosphopeptides; D000251:Adenosine Triphosphatases