Importance of p12 protein in Mason-Pfizer monkey virus assembly and infectivity.
Mason-Pfizer monkey virus (M-PMV) represents the prototype type D retrovirus, characterized by the assembly of intracytoplasmic A-type particles within the infected-cell cytoplasm. These immature particles migrate to the plasma membrane, where they are released by budding. The gag gene of M-PMV encodes a novel protein, p12, just 5' of the major capsid protein (CA) p27 on the polyprotein precursor. The function of p12 is not known, but an equivalent protein is found in mouse mammary tumor virus and is absent from the type C retroviruses. In order to determine whether the p12 protein plays a role in the intracytoplasmic assembly of capsids, a series of in-frame deletion mutations were constructed in the p12 coding domain. The mutant gag genes were expressed by a recombinant vaccinia virus-T7 polymerase-based system in CV-1 cells or in the context of the viral genome in COS-1 cells. In both of these high-level expression systems, mutant Gag precursors were competent to assemble but were not infectious. In contrast, when stable transfectant HeLa cell lines were established, assembly of the mutant precursors into capsids was drastically reduced. Instead, the polyprotein precursors remained predominantly soluble in the cytoplasm. These results show that while p12 is not required for the intracytoplasmic assembly of M-PMV capsids, under the conditions of low-level protein biosynthesis seen in virus-infected cells, it may assist in the stable association of polyprotein precursors for capsid assembly. Moreover, the presence of the p12 coding domain is absolutely required for the infectivity of M-PMV virions.
Sommerfelt
M A
MA
Department of Microbiology, University of Alabama, Birmingham 35294.
Rhee
S S
SS
Hunter
E
E
D000595:Amino Acid Sequence; D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D002460:Cell Line; D002522:Chlorocebus aethiops; D017353:Gene Deletion; D015683:Gene Products, gag / Q000235:genetics* / Q000378:metabolism; D015750:Genes, gag; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D016093:Mason-Pfizer monkey virus / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009024:Morphogenesis; D012516:Osteosarcoma; D014616:Vaccinia virus / Q000235:genetics; D014779:Virus Replication
D015683:Gene Products, gag; C078028:p12 protein, Mason-Pfizer monkey virus