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During human immunodeficiency virus type 1 (HIV-1) assembly, the primer tRNA for the reverse transcriptase-catalyzed synthesis of minus-strand strong-stop cDNA, tRNA3 Lys , is selectively packaged into the virus and annealed onto the primer binding site on the RNA genome. Annealing of tRNA3 Lys in HIV-1 is independent of polyprotein processing and is facilitated in vitro by p7 nucleocapsid (NCp7). We have previously shown that mutations in clusters of basic amino acids flanking the first Cys-His box in NC sequence inhibit annealing of tRNA3 Lys in vivo by 70 to 80%. In this report, we have investigated whether these NC mutations act through Pr55gag or Pr160gag-pol . In vivo placement of tRNA3 Lys is measured with total viral RNA as the source of primer tRNA-template in an in vitro reverse transcription assay. Cotransfection of COS cells with a plasmid coding for either mutant Pr55gag or mutant Pr160gag-pol , and with a plasmid containing HIV-1 proviral DNA, shows that only the NC mutations in Pr55gag inhibit tRNA3 Lys placement. The NC mutations in Pr55gag reduce viral infectivity by 95% and aretrans -dominant-negative, i.e., they inhibit genomic placement of tRNA3 Lys even in the presence of wild-type Pr55gag . This dominant phenotype may indicate that the mutant Pr55gag is disrupting an ordered Pr55gag structure responsible for the annealing of tRNA3 Lys to genomic RNA.

The Role of Pr55 gag in the Annealing of tRNA 3 Lys to Human Immunodeficiency Virus Type 1 Genomic RNA

The Role of Pr55 ^gag in the Annealing of tRNA ₃ ^Lys to Human Immunodeficiency Virus Type 1 Genomic RNA