Marked infidelity of human immunodeficiency virus type 1 reverse transcriptase at RNA and DNA template ends

Premal H. Patel, Bradley D. Preston

Research output: Contribution to journalArticlepeer-review

140 Scopus citations

Abstract

Human immunodeficiency virus type 1 (HIV-1) is genetically highly variable. This is attributed to the error-prone nature of HIV-1 replication and its proclivity for recombination. During replication and recombination, reverse transcriptase (RT) must polymerize DNA to the 5' ends of multiple RNA and DNA template termini while converting HIV-1 RNA to double-stranded DNA. We have determined the fidelity of HIV-1 RT in vitro during polymerization to the 5' ends of HIV-1 long terminal repeat DNA template sequences and to the end of a partial HIV-1 genomic RNA template that mimics a recombination intermediate. HIV-1 RT readily extended recessed DNA primers to form full- length blunt-end DNA·DNA and DNA·RNA duplexes. In addition, HIV-1 RT catalyzed high yields of products with one to four extra nucleotides at the 3' ends of the nascent DNAs. These products were formed processively via a nontemplated mechanism that is highly specific for the addition of purine nucleotides (A > G >> T ≥ C). Thus, HIV-1 RT is extremely unfaithful at both DNA and RNA template ends, introducing errors (extra nucleotides) in one out of every two or three nascent strands processively polymerized. This error rate is 1000 times higher than for HIV-1 RT-catalyzed errors at internal template positions. Blunt-end additions were also catalyzed by other retroviral RTs at relative rates of HIV-1 ≃ Moloney murine leukemia virus > avian myeloblastosis virus. These data suggest a potentially important mechanism for retroviral mutation mediated by nontemplated blunt-end addition of purines prior to forced copy-choice recombination.

Original languageEnglish (US)
Pages (from-to)549-553
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number2
DOIs
StatePublished - Jan 18 1994
Externally publishedYes

Keywords

  • DNA polymerase
  • mutagenesis
  • recombination
  • retrovirus

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'Marked infidelity of human immunodeficiency virus type 1 reverse transcriptase at RNA and DNA template ends'. Together they form a unique fingerprint.

Cite this