Characterization of recombinant murine leukemia virus integrase

Iris Dotan, Brian Scottoline, Timothy S. Heuer, Patrick O. Brown

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Retroviral integration involves two DNA substrates that play different roles. The viral DNA substrate is recognized by virtue of specific nucleotide sequences near the end of a double-stranded DNA molecule. The target DNA substrate is recognized at internal sites with little sequence preference; nucleosomal DNA appears to be preferred for this role. Despite this apparent asymmetry in the sequence, structure, and roles of the DNA substrates in the integration reaction, the existence of distinct binding sites for viral and target DNA substrates has been controversial. In this report, we describe the expression in Escherichia coli and purification of Moloney murine leukemia virus integrase as a fusion protein with glutathione S-transferase, characterization of its activity by using several model DNA substrates, and the initial kinetic characterization of its interactions with a model viral DNA substrate. We provide evidence for functionally and kinetically distinct binding sites for viral and target DNA substrates and describe a cross- linking assay for DNA binding at a site whose specificity is consistent with the target DNA binding site.

Original languageEnglish (US)
Pages (from-to)456-468
Number of pages13
JournalJournal of Virology
Volume69
Issue number1
StatePublished - Jan 1995
Externally publishedYes

Fingerprint

Murine leukemia virus
Integrases
Murine Leukemia Viruses
DNA
Viral DNA
Binding Sites
binding sites
Moloney murine leukemia virus
Glutathione Transferase
Escherichia coli
crosslinking
glutathione transferase

ASJC Scopus subject areas

  • Immunology

Cite this

Dotan, I., Scottoline, B., Heuer, T. S., & Brown, P. O. (1995). Characterization of recombinant murine leukemia virus integrase. Journal of Virology, 69(1), 456-468.

Characterization of recombinant murine leukemia virus integrase. / Dotan, Iris; Scottoline, Brian; Heuer, Timothy S.; Brown, Patrick O.

In: Journal of Virology, Vol. 69, No. 1, 01.1995, p. 456-468.

Research output: Contribution to journalArticle

Dotan, I, Scottoline, B, Heuer, TS & Brown, PO 1995, 'Characterization of recombinant murine leukemia virus integrase', Journal of Virology, vol. 69, no. 1, pp. 456-468.
Dotan, Iris ; Scottoline, Brian ; Heuer, Timothy S. ; Brown, Patrick O. / Characterization of recombinant murine leukemia virus integrase. In: Journal of Virology. 1995 ; Vol. 69, No. 1. pp. 456-468.
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