Functional domains of moloney murine leukemia virus integrase defined by mutation complementation analysis

Colleen Jonsson, George A. Donzella, Elena Gaucan, Christine M. Smith, Monica J. Roth

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Retroviral integrases perform two catalytic steps, 3′ processing and strand transfer, that result in the stable insertion of the retroviral DNA into the host genome. Mutant M-MuLV integrases were constructed to define the functional domains important for 3′ processing, strand transfer, and disintegration by in vitro assays. N-terminal mutants had no detectable 3′ processing activity, and only one mutant which lacks the HHCC domain, NΔ105, had strand transfer activity. Strand transfer mediated by NΔ105 showed preference for one site in the target DNA. Disintegration activity of N-terminal mutants decreased only minimally. In contrast, all C-terminal mutants truncated by more than 28 amino acids had no integration or disintegration activity. Activity on a single-strand disintegration substrate did not require a functional HHCC domain but did require most of the C-terminal region. Complementation analysis found that the HHCC region alone was able to function in trans to a promoter containing only the DD(35)E and C-terminal regions and to enhance integration site selection. Increasing the reducing conditions or adding the HHCC domain to NΔ105 reaction mixtures restored the wild-type strand transfer activity and range of target sites. The reducing agent affected Cys-209 in the DD(35)E region. The presence of C-209 was required for complementation of NΔ105 by the HHCC region.

Original languageEnglish (US)
Pages (from-to)4585-4597
Number of pages13
JournalJournal of Virology
Volume70
Issue number7
StatePublished - Dec 1 1996
Externally publishedYes

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Murine leukemia virus
Moloney murine leukemia virus
Integrases
mutation
mutants
Mutation
DNA
Reducing Agents
Genome
Amino Acids
reducing agents
promoter regions
amino acids
genome
assays

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Cite this

Functional domains of moloney murine leukemia virus integrase defined by mutation complementation analysis. / Jonsson, Colleen; Donzella, George A.; Gaucan, Elena; Smith, Christine M.; Roth, Monica J.

In: Journal of Virology, Vol. 70, No. 7, 01.12.1996, p. 4585-4597.

Research output: Contribution to journalArticle

Jonsson, C, Donzella, GA, Gaucan, E, Smith, CM & Roth, MJ 1996, 'Functional domains of moloney murine leukemia virus integrase defined by mutation complementation analysis', Journal of Virology, vol. 70, no. 7, pp. 4585-4597.
Jonsson, Colleen ; Donzella, George A. ; Gaucan, Elena ; Smith, Christine M. ; Roth, Monica J. / Functional domains of moloney murine leukemia virus integrase defined by mutation complementation analysis. In: Journal of Virology. 1996 ; Vol. 70, No. 7. pp. 4585-4597.
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