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Probing the Importance of tRNA Anticodon: Human Immunodeficiency Virus Type 1 (HIV-1) RNA Genome Complementarity with an HIV-1 That Selects tRNA^Glu for Replication

Department of Cell Biology,¹ Department of Microbiology,² Department of Molecular Genetics and Biochemistry, University of Alabama at Birmingham, Birmingham, Alabama 35294³

Received 23 December 2002/ Accepted 20 May 2003

The initiation of human immunodeficiency virus type 1 (HIV-1) reverse transcription occurs at the primer binding site (PBS) that is complementary to the 3'-terminal nucleotides of tRNA₃^Lys. Why all known strains of HIV-1 select tRNA₃^Lys for replication is unknown. Previous studies on the effect of altering the PBS of HIV-1 on replication identified an HIV-1 with a PBS complementary to tRNA^Glu. Since the virus was not initially designed to use tRNA^Glu, the virus had selected tRNA^Glu from the intracellular pool of tRNA for use in replication. Further characterization of HIV-1 that uses tRNA^Glu may provide new insights into the preference for tRNA₃^Lys. HIV-1 constructed with the PBS complementary to tRNA^Glu was more stable than HIV-1 with the PBS complementary to tRNA^Met or tRNA^His; however, all of these viruses eventually reverted back to using tRNA₃^Lys following growth in SupT1 cells or peripheral blood mononuclear cells (PBMCs). New HIV-1 mutants with nucleotides in U5 complementary to the anticodon of tRNA^Glu remained stable when grown in SupT1 cells or PBMCs, although the mutants grew more slowly than the wild-type virus. Sequence analysis of the U5 region and the PBS revealed additional mutations predicted to further promote tRNA-viral genome interaction. The results support the importance of the tRNA anticodon-genome interaction in the selection of the tRNA primer and highlight the fact that unique features of tRNA₃^Lys are exploited by HIV-1 for selection as the reverse transcription primer.

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