Reverse Transcriptase of Human Immunodeficiency Virus Can Use Either Human tRNALys 3or Escherichia coli tRNAGln 2as a Primer in an In vitro Primer-Utilization Assay

Although the reverse transcriptase (RT) of human immunodeficiency virus (HIV) uses human tRNALys 3as a primer of viral genome DNA synthesis in vivo, HIV RT binds Escherichia coli glutamine tRNA and in vitro-made human lysine tRNA with nearly equivalent affinities. We show that HIV RT can use either...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1992-10, Vol.89 (20), p.9652-9656
Main Authors: Kohlstaedt, Lori A., Steitz, Thomas A.
Format: Article
Language:English
Subjects:
Binding sites
Biochemistry
DNA
Gels
HIV
Nucleotides
Oligonucleotides
RNA
Transfer RNA
Viral genomes
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Summary:Although the reverse transcriptase (RT) of human immunodeficiency virus (HIV) uses human tRNALys 3as a primer of viral genome DNA synthesis in vivo, HIV RT binds Escherichia coli glutamine tRNA and in vitro-made human lysine tRNA with nearly equivalent affinities. We show that HIV RT can use either tRNALys 3or tRNAGln 2as a primer for DNA synthesis in vitro without the addition of any other host or viral proteins. E. coli tRNAGln 2can serve as a primer for HIV RT if a primer-binding site sequence complementary to the 3' end of tRNAGln 2is at the 3' end of the template. With this reduced template, the specificity of binding the proper tRNA is due to base-pairing between a bound tRNA to the primer-binding site of the viral RNA template rather than sequence-specific recognition of tRNALys 3by RT. If an 8-nucleotide viral sequence 3' to the primer-binding site is included in the template, then addition of Zn2+or Co2+is required for tRNALys 3-primed synthesis, and tRNAGln 2now fails to prime synthesis. The latter result implies that a template sequence adjacent to the primer-binding site and containing 6 nucleotides complementary to the anticodon loop of human tRNALys 3plays an active role in tRNA discrimination.
ISSN:0027-8424