Modulating target site selection during human immunodeficiency virus DNA integration in vitro with an engineered tethering factor

Details

Serval ID
serval:BIB_194316E543DF
Type
Article: article from journal or magazin.
Collection
Publications
Title
Modulating target site selection during human immunodeficiency virus DNA integration in vitro with an engineered tethering factor
Journal
Human Gene Therapy
Author(s)
Ciuffi  A., Diamond  T. L., Hwang  Y., Marshall  H. M., Bushman  F. D.
ISSN
1043-0342
Publication state
Published
Issued date
09/2006
Peer-reviewed
Oui
Volume
17
Number
9
Pages
960-7
Notes
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't --- Old month value: Sep
Abstract
The mechanisms controlling retroviral integration have been the topic of intense interest, in part because of adverse clinical events that occurred during retrovirus-mediated human gene therapy. Here we investigate the use of artificial tethering interactions to constrain retroviral integration site selection in an in vitro model. During normal infection, HIV DNA integration is favored in active cellular transcription units. One component of the targeting mechanism is the cellular LEDGF/p75 protein. LEDGF/p75 binds tightly to HIV integrase (IN) protein, and depletion of LEDGF/p75 from target cells results in reduced integration in transcription units, suggesting integration targeting by a tethering mechanism. We constructed and analyzed fusions of LEDGF/p75 or its IN-binding domain (IBD) to the DNA-binding domain of phage lambda repressor protein (lambdaR). In the presence of the lambdaR-LEDGF/p75 fusions, increased strand transfer by IN was seen in target DNA near lambdaR-binding sites in vitro . These data support the idea that a direct interaction between LEDGF/p75 and IN can mediate targeting via a tethering mechanism, and provide proof of concept for the idea that protein-protein interactions might be engineered to constrain integration site selection during human gene therapy.
Keywords
Base Sequence DNA Primers DNA, Viral/*genetics HIV/*genetics/physiology *Virus Integration
Pubmed
Web of science
Create date
22/02/2008 15:45
Last modification date
20/08/2019 13:50
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