Replication-defective Friend murine leukemia virus particles containing uncleaved gag polyproteins and decreased levels of envelope glycoprotein.
An erythroleukemia cell clone, 7C, which failed to produce reverse transcriptase-containing virions or infectious virus, was found to produce noninfectious virus particles by gradient banding of [3H]leucine- and [3H]uridine-labeled virions. The RNA from the 7C virus was shown to consist of the normal 70S size component, which converted to 35S upon heat denaturation. In contrast, the 7C virion proteins showed multiple defects. Analysis of the virion proteins by gel electrophoresis demonstrated that the pr65 gag precursor was incorporated into the 7C virus and that the processing of this precursor was severely diminished. Polymerase proteins pr180gag-pol and pr120pol were also detected in virions, and a third possible polymerase protein, p70, was reduced in size compared to its normal counterpart, p80. Incorporation of the viral gp70 glycoprotein into particles was also reduced 10-fold, despite synthesis and incorporation of gp70 into the 7C cell membrane in normal amounts. Pulse-chase analysis of the synthesis of the viral gag and env proteins in 7C cells showed greatly reduced amounts of pr180gag-pol, pr65gag, p80gag, and p42gag, whereas pr90env, gp70, and spleen focus-forming virus-specific gp55 were synthesized and processed normally. These results suggested that at least one defect in 7C virus was impaired cleavage of gag or pol proteins or both, most likely due to a lack of the appropriate viral protease, and that this lack of cleavage might affect incorporation of gp70 into virus particles.
D000818:Animals; D002478:Cells, Cultured; D002999:Clone Cells / Q000382:microbiology; D003673:Defective Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D005622:Friend murine leukemia virus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015683:Gene Products, gag; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D009374:Neoplasms, Experimental; D011498:Protein Precursors / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D015683:Gene Products, gag; D011498:Protein Precursors; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase