Identification and characterization of the herpesvirus saimiri oncoprotein STP-C488.
The protein encoded by herpesvirus saimiri transforming gene STP-C488 was identified and characterized. Antibodies were produced in rabbits by immunization with keyhole limpet hemocyanin-conjugated synthetic peptides specific for the predicted sequence of STP-C488. STP-C488-encoded protein was detected in recombinant Escherichia coli, transformed Rat-1 cells, transfected COS-1 cells, and in common marmoset T lymphocytes immortalized by herpesvirus saimiri strain 488. STP-C488 protein was sensitive to treatment by bacterial collagenase, consistent with the 18 uninterrupted collagenlike repeats predicted by the DNA sequence. The apparent molecular size of STP-C488 in sodium dodecyl sulfate (SDS)-polyacrylamide gels (20 to 22 kDa) was considerably larger than that predicted from the DNA sequence (9.9 kDa). Using indirect immunofluorescence tests and subcellular fractionation, STP-C488 was found to be membrane bound, primarily in perinuclear compartments. The 18 uninterrupted collagenlike repeats, sensitivity to collagenase, location in the cell, and anomalous migration through SDS-polyacrylamide gels suggest an unusual, membrane-associated, fibrous structure for this transforming herpesvirus oncoprotein.
Jung
J U
JU
New England Regional Primate Research Center, Harvard Medical School, Southborough, Massachusetts 01772.
Desrosiers
R C
RC
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002460:Cell Line; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005814:Genes, Viral; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics*; D003012:Microbial Collagenase; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009856:Oncogene Proteins, Viral / Q000032:analysis / Q000235:genetics*; D016366:Open Reading Frames; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010957:Plasmids; D014162:Transfection; D014771:Virion / Q000235:genetics / Q000378:metabolism
D000906:Antibodies; D004279:DNA, Viral; D008565:Membrane Proteins; D009856:Oncogene Proteins, Viral; D010455:Peptides; D003012:Microbial Collagenase