       Document 0110
 DOCN  M9630110
 TI    Directed integration of viral DNA mediated by fusion proteins consisting
       of human immunodeficiency virus type 1 integrase and Escherichia coli
       LexA protein.
 DT    9603
 AU    Goulaouic H; Chow SA; Department of Molecular and Medical Pharmacology,
       UCLA School of; Medicine 90095, USA.
 SO    J Virol. 1996 Jan;70(1):37-46. Unique Identifier : AIDSLINE MED/96099411
 AB    We tested whether the selection of target sites can be manipulated by
       fusing retroviral integrase with a sequence-specific DNA-binding
       protein. A hybrid protein that has the Escherichia coli LexA protein
       fused to the C terminus of the human immunodeficiency virus type 1
       integrase was constructed. The fusion protein, IN1-288/LA, retained the
       catalytic activities in vitro of the wild-type human immunodeficiency
       virus type 1 integrase (WT IN). Using an in vitro integration assay that
       included multiple DNA fragment as the target DNA, we found that
       IN1-288/LA preferentially integrated viral DNA into the fragment
       containing a DNA sequence specifically bound by LexA protein. No bias
       was observed when the LexA-binding sequence was absent, when the fusion
       protein was replaced by WT IN, or when LexA protein was added in the
       reaction containing IN1-288/LA. A majority of the integration events
       mediated by IN1-288/LA occurred within 30 bp of DNA flanking the
       LexA-binding sequence. The specificity toward the LexA-binding sequence
       and the distribution and frequency of target site usage were unchanged
       when the integrase component of the fusion protein was replaced with a
       variant containing a truncation at the N or C terminus or both,
       suggesting that the domain involved in target site selection resides in
       the central core region of integrase. The integration bias observed with
       the integrase-LexA hybrid shows that one effective means of altering the
       selection of DNA sites for integration is by fusing integrase to a
       sequence-specific DNA-binding protein.
 DE    Bacterial Proteins/GENETICS/*METABOLISM  Base Sequence  Binding Sites
       DNA Nucleotidyltransferases/GENETICS/*METABOLISM  DNA-Binding
       Proteins/GENETICS/*METABOLISM  DNA, Viral/METABOLISM  Escherichia
       coli/*METABOLISM  Feasibility Studies  Human  HIV-1/*ENZYMOLOGY
       Molecular Sequence Data  Recombinant Fusion Proteins/GENETICS/METABOLISM
       Substrate Specificity  Support, Non-U.S. Gov't  Support, U.S. Gov't,
       Non-P.H.S.  Support, U.S. Gov't, P.H.S.  Virus Integration/*GENETICS
       JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

