Activities of the feline immunodeficiency virus integrase protein produced in Escherichia coli.

Abstract:

:Retroviral DNA integration requires the activity of at least one viral protein, the integrase (IN) protein. We cloned and expressed the integrase gene of feline immunodeficiency virus (FIV) in Escherichia coli as a fusion to the malE gene and purified the IN fusion protein by affinity chromatography. The protein is active in site-specific cleavage of the viral DNA ends, DNA strand transfer, and disintegration. FIV IN has a relaxed viral DNA substrate requirement: it cleaves and integrates FIV DNA termini, human immunodeficiency virus DNA ends, and Moloney murine leukemia virus DNA ends with high efficiencies. In the cleavage reaction, IN exposes a specific phosphodiester bond near the viral DNA end to nucleophilic attack. In vitro, either H2O, glycerol, or the 3' OH group of the viral DNA terminus can serve as nucleophile in this reaction. We found that FIV IN preferentially uses the 3' OH ends of the viral DNA as nucleophile, whereas HIV IN protein preferentially uses H2O and glycerol as nucleophiles.

journal_name

J Virol

journal_title

Journal of virology

authors

Vink C,van der Linden KH,Plasterk RH

doi

10.1128/JVI.68.3.1468-1474.1994

subject

Has Abstract

pub_date

1994-03-01 00:00:00

pages

1468-74

issue

3

eissn

0022-538X

issn

1098-5514

journal_volume

68

pub_type

杂志文章