Lens epithelium-derived growth factor fusion proteins redirect HIV-1 DNA integration

Ferris, Andrea L.; Wu, Xiaolin; Hughes, Christina; Stewart, Claudia; Smith, Steven J.; Milne, Thomas A.; Wang, Gang Greg; Shun, Ming Chieh; Allis, C. David; Engelman, Alan; Hughes, Stephen H.

doi:10.1073/pnas.0914142107

Cited by 127 publications

(138 citation statements)

References 34 publications

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“…The consequence of LEDGF tethering is that HIV-1 reverse transcripts typically integrate within active transcription units (36). Steve Hughes and collaborators Alan Engelman (Dana-Farber Institute) and David Allis (Rockefeller University) have fused the integrase-binding domain of LEDGF to chromatin-binding domains of other proteins (37) to test the idea that the HIV-1 target site bias is determined primarily by the specificity of the chromatinbinding domain of the host protein with which HIV-1 integrase interacts. In his talk, Hughes discussed swapping experiments involving the plant homeodomain PHD finger motif of the histone demethylase JARID1A, the PHD finger and atypical bromodomain region of the histone methyltransferase MLL, and the chromodomain from polycomb group protein Cbx7.…”

Section: Targeting Specificitymentioning

confidence: 99%

Telomerase and retrotransposons: Reverse transcriptases that shaped genomes

Belfort

Curcio

Lue

2011

Proc. Natl. Acad. Sci. U.S.A.

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Section: Targeting Specificitymentioning

confidence: 99%

Telomerase and retrotransposons: Reverse transcriptases that shaped genomes

Belfort

Curcio

Lue

2011

Proc. Natl. Acad. Sci. U.S.A.

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“…These observations have suggested that different cellularbinding partners of retroviral integrases are likely to be responsible for integration target-site selection. However, to date, only one example has been reported: lens epithelium-derived growth factor (LEDGF/p75), which functions as a bimodal tether that engages HIV-1 intasomes and navigates them to active genes (8)(9)(10)(11)(12)(13)(14). Cellular cofactors of other retroviral genera are currently unknown.…”

mentioning

confidence: 99%

BET proteins promote efficient murine leukemia virus integration at transcription start sites

Sharma¹,

Larue²,

Plumb³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

185

267

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The selection of chromosomal targets for retroviral integration varies markedly, tracking with the genus of the retrovirus, suggestive of targeting by binding to cellular factors. γ-Retroviral murine leukemia virus (MLV) DNA integration into the host genome is favored at transcription start sites, but the underlying mechanism for this preference is unknown. Here, we have identified bromodomain and extraterminal domain (BET) proteins (Brd2, -3, -4) as cellular-binding partners of MLV integrase. We show that purified recombinant Brd4(1-720) binds with high affinity to MLV integrase and stimulates correct concerted integration in vitro. JQ-1, a small molecule that selectively inhibits interactions of BET proteins with modified histone sites impaired MLV but not HIV-1 integration in infected cells. Comparison of the distribution of BET protein-binding sites analyzed using ChIP-Seq data and MLV-integration sites revealed significant positive correlations. Antagonism of BET proteins, via JQ-1 treatment or RNA interference, reduced MLV-integration frequencies at transcription start sites. These findings elucidate the importance of BET proteins for MLV integration efficiency and targeting and provide a route to developing safer MLV-based vectors for human gene therapy. (1-4). The selection of chromosomal targets for retroviral integration varies markedly, tracking with the genus of the retrovirus studied (5-7). For example, the γ-retroviruses favor integration near transcription start sites, whereas lentiviruses favor integration within transcription units. These observations have suggested that different cellularbinding partners of retroviral integrases are likely to be responsible for integration target-site selection. However, to date, only one example has been reported: lens epithelium-derived growth factor (LEDGF/p75), which functions as a bimodal tether that engages HIV-1 intasomes and navigates them to active genes (8-14). Cellular cofactors of other retroviral genera are currently unknown.The molecular mechanisms of γ-retroviral murine leukemia virus (MLV) integration are of particular significance because MLV-based vectors are used for human gene therapy. In clinical trials, the use of γ-retroviral vectors to correct primary immunodeficiencies has been curative, but adverse events have occurred associated with insertion of MLV-based vectors near protooncogenes (reviewed in refs. 15-18). The identification of cellular factors for γ-retroviruses may provide mechanistic clues to facilitate the development of safer gene-therapy vectors.In this report, we have identified the bromodomain and extraterminal domain (BET) proteins (Brd2, -3, -4) as the cellularbinding partners of MLV IN and demonstrate their significance for stimulating and targeting MLV integration at transcription start sites. (Table 1, Table S1, and Fig. S1). Of these, Brd4 and Brd3 were the top hits in NIH 3T3 and Sup-T1 cells, respectively. Differential pull-down levels of these proteins (Table 1) could be attributable to the varying expression le...

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“…However, integrase and the IBD can be mutated in parallel so that they still bind one another, but integrase no longer binds endogenous LEDGF (11). In this study Ferris et al (5) show that the half of LEDGF that binds to chromatin can be swapped to essentially any CBD of desired specificity. Although the extent to which retroviral target specificity can be directed remains to be determined, manipulating the dual-anchor LEDGF, which bridges the PIC and chromatin before integration, is the most promising strategy to date.…”

mentioning

confidence: 95%

“…Regardless of the functional significance of target-site selection for the virus, the study by Ferris et al (5) shows that targeting can be redirected by substituting the CBD of the LEDGF with a different anchor. This finding has potential importance in the application of retroviral vectors for gene therapy.…”

mentioning

confidence: 99%

“…The study by Ferris et al (5) tests this hypothesis by replacing the PWWP and AT hooks of LEDGF with other CBDs with known specificity (Fig. 1): the plant homeodomain finger from inhibitor of growth protein 2 (ING2), which binds histone 3 lysine 4 trimethylations (H3K4me3), and the chromodomain of heterochromatin protein 1-α (HP1α), which binds histone 3 that is methylated on lysine 9 (H3K9me2,3).…”

mentioning

confidence: 99%

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