The impact of molecular manipulation in residue 114 of human immunodeficiency virus type-1 reverse transcriptase on dNTP substrate binding and viral replication

Sarah K. Van Cor-Hosmer, Waaqo Daddacha, Z. Kelly, Amy Tsurumi, Edward M. Kennedy, Baek Kim

Research output: Contribution to journalArticle

Abstract

Human immunodeficiency virus type-1 (HIV-1) reverse transcriptase (RT) has a unique tight binding to dNTP substrates. Structural modeling of Ala-114 of HIV-1 RT suggests that longer side chains at this residue can reduce the space normally occupied by the sugar moiety of an incoming dNTP. Indeed, mutations at Ala-114 decrease the ability of RT to synthesize DNA at low dNTP concentrations and reduce the dNTP-binding affinity (K d) of RT. However, the K d values of WT and A114C RT remained equivalent with an acyclic dNTP substrate. Finally, mutant A114 RT HIV-1 vectors displayed a greatly reduced transduction in nondividing human lung fibroblasts (HLFs), while WT HIV-1 vector efficiently transduced both dividing and nondividing HLFs. Together these data support that the A114 residue of HIV-1 RT plays a key mechanistic role in the dNTP binding of HIV-1 RT and the unique viral infectivity of target cell types with low dNTP pools.

Original languageEnglish (US)
Pages (from-to)393-401
Number of pages9
JournalVirology
Volume422
Issue number2
DOIs
StatePublished - Jan 20 2012
Externally publishedYes

Fingerprint

HIV-1
RNA-Directed DNA Polymerase
Fibroblasts
Lung
Human immunodeficiency virus 1 reverse transcriptase
Mutation
DNA

Keywords

  • Cellular dNTP pools
  • DNTP binding affinity
  • HIV-1
  • Reverse transcriptase
  • Steady and presteady kinetics

ASJC Scopus subject areas

  • Virology

Cite this

The impact of molecular manipulation in residue 114 of human immunodeficiency virus type-1 reverse transcriptase on dNTP substrate binding and viral replication. / Van Cor-Hosmer, Sarah K.; Daddacha, Waaqo; Kelly, Z.; Tsurumi, Amy; Kennedy, Edward M.; Kim, Baek.

In: Virology, Vol. 422, No. 2, 20.01.2012, p. 393-401.

Research output: Contribution to journalArticle

Van Cor-Hosmer, Sarah K. ; Daddacha, Waaqo ; Kelly, Z. ; Tsurumi, Amy ; Kennedy, Edward M. ; Kim, Baek. / The impact of molecular manipulation in residue 114 of human immunodeficiency virus type-1 reverse transcriptase on dNTP substrate binding and viral replication. In: Virology. 2012 ; Vol. 422, No. 2. pp. 393-401.
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