Maturation of dimeric viral RNA of Moloney murine leukemia virus.
We have analyzed the dimeric RNA present in Moloney murine leukemia virus (MoMuLV) particles. We found that the RNA in newly released virions is in a conformation different from that in mature virions, since it has a different electrophoretic mobility in nondenaturing agarose gels and dissociates into monomers at a lower temperature. On the basis of these results, we suggest that the RNA initially packaged into nascent virions is already dimeric but that the dimer undergoes a maturation process after the virus is released from the cell. In further experiments, we tested the possibility that this maturation event is linked to the maturation cleavage of the virion proteins, which is catalyzed by the viral protease (PR). We found that the dimeric RNA isolated from PR- mutant virions resembles that from immature virions: it has a lower electrophoretic mobility and a lower sedimentation rate, and it also dissociates at a lower temperature than does RNA from mature wild-type virions. When Kirsten sarcoma virus is rescued by a PR- mutant or by a somewhat leaky cysteine array mutant of MoMuLV, its RNA also exhibits a electrophoretic mobility lower than that in the wild-type pseudotype. These results suggest that the maturation of dimeric RNA in released virus particles requires the cleavage of the Gag precursor and the presence of an intact cysteine array in the released nucleocapsid protein.
Fu
W
W
Laboratory of Molecular Virology and Carcinogenesis, NCI-Frederick Cancer Research and Development Center, Maryland 21702.
Rein
A
A
D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004279:DNA, Viral / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D046911:Macromolecular Substances; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000378:metabolism*; D009691:Nucleic Acid Denaturation; D012330:RNA, Double-Stranded / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013816:Thermodynamics; D014529:Uridine / Q000378:metabolism; D014771:Virion / Q000235:genetics / Q000378:metabolism
D004279:DNA, Viral; D046911:Macromolecular Substances; D012330:RNA, Double-Stranded; D014529:Uridine