Truncating α-Helix E′ of p66 human immunodeficiency virus reverse transcriptase modulates RNase H function and impairs DNA strand transfer

Madhumita Ghosh, Kathryn J. Howard, Craig Eugene Cameron, Stephen Benkovic, Stephen H. Hughes, Stuart F.J. Le Grice

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

The properties of recombinant p66/p51 human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) containing C-terminal truncations in its p66 polypeptide were evaluated. Deletion end points partly or completely removed α-helix E′ of the RNase H domain (p66Δ8/p51 and p66Δ16/p51, respectively), while mutant p66Δ23/p51 lacked αE′ and the β5′-αE′ connecting loop. Although dimerization and DNA polymerase properties of all mutants were not significantly different from those of the parental enzyme, p66Δ16/p51 and p66Δ23/ p51 RT lacked ribonuclease H (RNase H) activity. In contrast, RT mutant p66Δ8/p51 retained endonuclease activity but lacked the directional processing feature of the parental enzyme. Despite retaining full endoribonuclease function, p66Δ8/p51 RT barely supported transfer of nascent (-)-strand DNA between RNA templates representing the 5′ and 3′ ends of retroviral genome, shedding light on the requirement for the endonuclease and directional processing functions of the RNase H domain during replication.

Original languageEnglish (US)
Pages (from-to)7068-7076
Number of pages9
JournalJournal of Biological Chemistry
Volume270
Issue number13
DOIs
StatePublished - Mar 31 1995

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Ribonuclease H
HIV Reverse Transcriptase
Endonucleases
DNA
Endoribonucleases
Dimerization
DNA-Directed DNA Polymerase
Enzymes
Processing
HIV-1
Genes
Genome
RNA
Peptides

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ghosh, Madhumita ; Howard, Kathryn J. ; Cameron, Craig Eugene ; Benkovic, Stephen ; Hughes, Stephen H. ; Le Grice, Stuart F.J. / Truncating α-Helix E′ of p66 human immunodeficiency virus reverse transcriptase modulates RNase H function and impairs DNA strand transfer. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 13. pp. 7068-7076.
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abstract = "The properties of recombinant p66/p51 human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) containing C-terminal truncations in its p66 polypeptide were evaluated. Deletion end points partly or completely removed α-helix E′ of the RNase H domain (p66Δ8/p51 and p66Δ16/p51, respectively), while mutant p66Δ23/p51 lacked αE′ and the β5′-αE′ connecting loop. Although dimerization and DNA polymerase properties of all mutants were not significantly different from those of the parental enzyme, p66Δ16/p51 and p66Δ23/ p51 RT lacked ribonuclease H (RNase H) activity. In contrast, RT mutant p66Δ8/p51 retained endonuclease activity but lacked the directional processing feature of the parental enzyme. Despite retaining full endoribonuclease function, p66Δ8/p51 RT barely supported transfer of nascent (-)-strand DNA between RNA templates representing the 5′ and 3′ ends of retroviral genome, shedding light on the requirement for the endonuclease and directional processing functions of the RNase H domain during replication.",
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Truncating α-Helix E′ of p66 human immunodeficiency virus reverse transcriptase modulates RNase H function and impairs DNA strand transfer. / Ghosh, Madhumita; Howard, Kathryn J.; Cameron, Craig Eugene; Benkovic, Stephen; Hughes, Stephen H.; Le Grice, Stuart F.J.

In: Journal of Biological Chemistry, Vol. 270, No. 13, 31.03.1995, p. 7068-7076.

Research output: Contribution to journalArticle

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AU - Ghosh, Madhumita

AU - Howard, Kathryn J.

AU - Cameron, Craig Eugene

AU - Benkovic, Stephen

AU - Hughes, Stephen H.

AU - Le Grice, Stuart F.J.

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