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M9620233.TXT
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1996-02-26
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Document 0233
DOCN M9620233
TI In vitro integration of human immunodeficiency virus type 1 cDNA into
targets containing protein-induced bends.
DT 9602
AU Bor YC; Bushman FD; Orgel LE; Salk Institute for Biological Studies, San
Diego, CA 92186-8500,; USA.
SO Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10334-8. Unique Identifier
: AIDSLINE MED/96036079
AB Integration of human immunodeficiency virus type 1 cDNA into a target
DNA can be strongly influenced by the conformation of the target. For
example, integration in vitro is sometimes favored in target DNAs
containing sequence-directed bends or DNA distortions caused by bound
proteins. We have analyzed the effect of DNA bending by studying
integration into two well-characterized protein-DNA complexes:
Escherichia coli integration host factor (IHF) protein bound to a phage
IHF site, and the DNA binding domain of human lymphoid enhancer factor
(LEF) bound to a LEF site. Both of these proteins have previously been
reported to bend DNA by approximately 140 degrees. Binding of IHF
greatly increases the efficiency of in vitro integration at hotspots
within the IHF site. We analyzed a series of mutants in which the IHF
site was modified at the most prominent hotspot. We found that each
variant still displayed enhanced integration upon IHF binding. Evidently
the local sequence is not critical for formation of an IHF hotspot. LEF
binding did not create preferred sites for integration. The different
effects of IHF and LEF binding can be rationalized in terms of the
different proposed conformations of the two protein-DNA complexes.
DE Bacterial Proteins/CHEMISTRY/*METABOLISM Base Sequence Comparative
Study DNA Nucleotidyltransferases/BIOSYNTHESIS/*METABOLISM DNA Primers
DNA-Binding Proteins/METABOLISM DNA,
Complementary/CHEMISTRY/*METABOLISM DNA, Viral/CHEMISTRY/*METABOLISM
Human HIV-1/*GENETICS/METABOLISM Models, Structural Molecular
Sequence Data Nucleic Acid Conformation Polymerase Chain Reaction
Protein Conformation Recombinant
Proteins/BIOSYNTHESIS/CHEMISTRY/METABOLISM Support, U.S. Gov't, P.H.S.
*Virus Integration JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).