trans-acting inhibition of genomic RNA dimerization by Rous sarcoma virus matrix mutants.
The genomic RNA of retroviruses exists within the virion as a noncovalently linked dimer. Previously, we identified a mutant of the viral matrix (MA) protein of Rous sarcoma virus that disrupts viral RNA dimerization. This mutant, Myr1E, is modified at the N terminus of MA by the addition of 10 amino acids from the Src protein, resulting in the production of particles containing monomeric RNA. Dimerization is reestablished by a single amino acid substitution that abolishes myristylation (Myr1E-). To distinguish between cis and trans effects involving Myr1E, additional mutations were generated. In Myr1E.cc and Myr1E-.cc, different nucleotides were utilized to encode the same protein as Myr1E and Myr1E-, respectively. The alterations in RNA sequence did not change the properties of the viral mutants. Myr1E.ATG- was constructed so that translation began at the gag AUG, resulting in synthesis of the wild-type Gag protein but maintenance of the src RNA sequence. This mutant had normal infectivity and dimeric RNA, indicating that the src sequence did not prevent dimer formation. All of the src-containing RNA sequences formed dimers in vitro. Examination of MA-green fluorescent protein fusion proteins revealed that the wild-type and mutant MA proteins Myr1E.ATG-, Myr1E-, and Myr1E-.cc had distinctly different patterns of subcellular localization compared with Myr1E and Myr1E.cc MA proteins. This finding suggests that proper localization of the MA protein may be required for RNA dimer formation and infectivity. Taken together, these results provide compelling evidence that the genomic RNA dimerization defect is due to a trans-acting effect of the mutant MA proteins.
Garbitt
R A
RA
Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Milton S. Hershey Medical Center, Hershey, Pennsylvania 17033, USA.
Albert
J A
JA
Kessler
M D
MD
Parent
L J
LJ
D000483:Alleles; D000595:Amino Acid Sequence; D001358:Avian Sarcoma Viruses / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D019281:Dimerization; D015750:Genes, gag; D008969:Molecular Sequence Data; D009154:Mutation; D011533:Proviruses / Q000472:pathogenicity; D012367:RNA, Viral / Q000737:chemistry*; D014763:Viral Matrix Proteins / Q000032:analysis / Q000502:physiology*
D012367:RNA, Viral; D014763:Viral Matrix Proteins