Influence of substrate structure on disintegration activity of Moloney murine leukemia virus integrase

G. A. Donzella, Colleen Jonsson, M. J. Roth

Research output: Contribution to journalArticle

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Abstract

The disintegration activity of Moloney murine leukemia virus (M-MuLV) integrase (IN) was investigated through structural and sequence modifications of a Y substrate that resembles an integration intermediate. The Y substrates, constructed from individual oligonucleotides, contain a single viral long terminal repeat (LTR) joined to a nicked target DNA. Truncation of the double-stranded LTR sequences distal to the conserved 5'-CA-3' dinucleotide progressively diminished disintegration activity. M-MuLV IN was also able to catalyze disintegration of a heterologous double-stranded LTR sequence. Significantly, the activity of M-MuLV IN on single-stranded LTR Y substrates was more dependent on the sequence and length of the LTR strand than that reported for human immunodeficiency virus type 1 (HIV-1) IN. Modifications introduced at the Y-substrate junction demonstrated that the 3'-hydroxyl group at the terminus of the target strand was necessary for efficient joining of the target DNA strands. The presence of a 2'-hydroxyl group at the 3' end of the target strand, as well as a single-nucleotide gap at the LTR-target junction, reduced disintegration activity. The absence of hydroxyl groups on the terminal nucleotide abolished joining of the target strands. The results presented here suggest that M-MuLV IN disintegration activity is dependent on substantially different LTR sequence requirements than those reported for HIV-1 IN and may be mediated primarily through a structural recognition event.

Original languageEnglish (US)
Pages (from-to)7077-7087
Number of pages11
JournalJournal of Virology
Volume67
Issue number12
StatePublished - Jan 1 1993
Externally publishedYes

Fingerprint

Murine leukemia virus
Moloney murine leukemia virus
Integrases
terminal repeat sequences
Terminal Repeat Sequences
Hydroxyl Radical
Human immunodeficiency virus 1
HIV-1
Nucleotides
nucleotides
DNA
oligonucleotides
Oligonucleotides

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Cite this

Influence of substrate structure on disintegration activity of Moloney murine leukemia virus integrase. / Donzella, G. A.; Jonsson, Colleen; Roth, M. J.

In: Journal of Virology, Vol. 67, No. 12, 01.01.1993, p. 7077-7087.

Research output: Contribution to journalArticle

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