LEDGF/p75 Proteins with Alternative Chromatin Tethers Are Functional HIV-1 Cofactors

Meehan, Anne M.; Saenz, Dyana T.; Morrison, James H.; Garcia-Rivera, Jose A.; Peretz, Mary; Llano, Manuel; Poeschla, Eric M.

doi:10.1371/journal.ppat.1000522

Cited by 61 publications

(91 citation statements)

References 70 publications

(154 reference statements)

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“…Replacing the N-terminal end of LEDGF/p75, containing all chromatin binding elements, with full-length HDGF resulted in similar staining (Fig. 3, g and n, respectively), demonstrating that the N-terminal domain of LEDGF/p75 can be replaced with alternative chromatin binding elements, in line with earlier reports (17,19).…”

Section: Generation Of Ledgf/p75supporting

confidence: 89%

“…3) (19,36,37,68), and loss of interaction with mitotic chromatin (19,37). The PWWP domain of LEDGF/p75 appears to exert a dominant effect on the protein intranuclear localization, excluding it from the nucleoli (19,46,68,74).…”

Section: Discussionmentioning

confidence: 99%

“…Cells depleted for LEDGF/p75 (13,14) or somatic knock-out cells (15,16) inhibit productive HIV infection. In addition, fusion proteins in which the LEDGF/p75 DNA binding portion is replaced with an alternative chromatin binding domain efficiently tether the PIC to the host cell chromatin and support lentiviral vector transduction (17)(18)(19)(20). We and others demonstrated that these fusion proteins retarget proviral integration to the regions bound by the specific chromatin binding domain (17,18).…”

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confidence: 95%

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Role of the PWWP Domain of Lens Epithelium-derived Growth Factor (LEDGF)/p75 Cofactor in Lentiviral Integration Targeting

Gijsbers

Vets

Rijck

et al. 2011

Journal of Biological Chemistry

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LEDGF/p75 is a chromatin-interacting, cellular cofactor of HIV integrase that dictates lentiviral integration site preference. In this study we determined the role of the PWWP domain of LEDGF/p75 in tethering and targeting of the lentiviral pre-integration complex, employing potent knockdown cell lines allowing analysis in the absence of endogenous LEDGF/p75. Deletion of the PWWP domain resulted in a diffuse subnuclear distribution pattern, loss of interaction with condensed chromatin, and failure to rescue proviral integration, integration site distribution, and productive virus replication. Substitution of the PWWP domain of LEDGF/p75 with that of hepatoma-derived growth factor or HDGF-related protein-2 rescued viral replication and lentiviral integration site distribution in LEDGF/p75-depleted cells. Replacing all chromatin binding elements of LEDGF/p75 with full-length hepatoma-derived growth factor resulted in more integration in genes combined with a preference for CpG islands. In addition, we showed that any PWWP domain targets SMYD1-like sequences. Analysis of integration preferences of lentiviral vectors for epigenetic marks indicates that the PWWP domain is critical for interactions specifying the relationship of integration sites to regions enriched in specific histone post-translational modifications.Stable integration of the viral DNA into the host genome is one of the hallmarks of retroviral replication and has profound consequences for both the virus and the host. For the virus it is essential to direct integration in the host cell chromatin to sites that allow efficient gene expression to fulfill a successful infection cycle. Retroviruses from different genera have evolved to integrate their genomic DNA at different sites in the genome of their respective hosts. Human immunodeficiency virus (HIV) and other lentiviruses have a strong preference for integration in active transcription units (1), and murine leukemia virus (MLV) genomes preferentially integrate near transcription start sites and CpG island regions (2), whereas avian sarcoma leukosis virus (3, 4) and the human T-cell leukemia virus display a much weaker preference for these features (5, 6), only weakly favoring integration in active transcription units. Deciphering the mechanisms that dictate integration site selection is instrumental to better understand basic retrovirology and its clinical applications such as drug development and gene therapy.Studies of yeast retrotransposons demonstrated that cellular cofactors determine the integration site preference (7-9). Likewise, interactions between HIV integrase (IN) 4 and the host cell protein lens epithelium-derived growth factor (LEDGF/p75) (10) direct HIV integration targeting. LEDGF/p75 specifically binds IN via its integrase binding domain (LEDGF ) ( Fig. 1) and functions as a molecular tether during lentiviral integration, bridging the viral pre-integration complex (PIC) with the host cell chromatin (11-13). Cells depleted for LEDGF/p75 (13, 14) or somatic knock-out cells (15, 1...

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Section: Generation Of Ledgf/p75supporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 95%

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Role of the PWWP Domain of Lens Epithelium-derived Growth Factor (LEDGF)/p75 Cofactor in Lentiviral Integration Targeting

Gijsbers

Vets

Rijck

et al. 2011

Journal of Biological Chemistry

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“…If the PWWP domain of LEDGF is removed, the truncated protein cannot support HIV-1 replication. However, fusion proteins in which the PWWP domain of LEDGF is replaced with the linker histone H1 or with a peptide derived from the chromatin binding Kaposi's sarcoma herpes virus LANA protein do support efficient HIV-1 replication (23). This result means that these fusion proteins are able to bind both chromatin and the PIC in a way that supports viral integration.…”

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confidence: 99%

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

Ferris

Hughes

et al. 2010

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

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Lens epithelium-derived growth factor (LEDGF) fusion proteins can direct HIV-1 DNA integration to novel sites in the host genome. The C terminus of LEDGF contains an integrase binding domain (IBD), and the N terminus binds chromatin. LEDGF normally directs integrations to the bodies of expressed genes. Replacing the N terminus of LEDGF with chromatin binding domains (CBDs) from other proteins changes the specificity of HIV-1 DNA integration. We chose two well-characterized CBDs: the plant homeodomain (PHD) finger from ING2 and the chromodomain from heterochromatin binding protein 1α (HP1α). The ING2 PHD finger binds H3K4me3, a histone mark that is associated with the transcriptional start sites of expressed genes. The HP1α chromodomain binds H3K9me2,3, histone marks that are widely distributed throughout the genome. A fusion protein in which the ING2 PHD finger was linked to the LEDGF IBD directed integrations near the start sites of expressed genes. A similar fusion protein in which the HP1α chromodomain was linked to the LEDGF IBD directed integrations to sites that differed from both the PHD finger fusion-directed and LEDGF-directed integration sites. The ability to redirect HIV-1 DNA integration may help solve the problems associated with the activation of oncogenes when retroviruses are used in gene therapy.chromatin | histone marks | lentiviral vectors R etroviruses can integrate their DNAs at many sites in host DNA (1), but different retroviruses have different integration site preferences (2-9). HIV-1 and simian immunodeficiency virus DNAs preferentially integrate into expressed genes, murine leukemia virus (MLV) DNA preferentially integrates near transcriptional start sites (TSSs), and avian sarcoma leukosis virus (ASLV) and human T cell leukemia virus (HTLV) DNAs integrate nearly randomly, showing a slight preference for genes. This suggests that the preintegration complexes (PICs) of the different retroviruses interact with different factors in host cell chromatin and that the integration site preferences are determined by the distribution of the different host cell factors. Studies of the specificity of integration of yeast retrotransposons, which are distantly related to retroviruses, provide strong support for this interpretation (10-12). The host factors that MLV, ASLV, and HLTV PICs interact with are not known.

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“…Importantly, a viable mouse embryo fibroblast LEDGF knockout (MEF-KO) cell line exists that supports HIV-1 replication, albeit at a reduced level, enabling the potential role of this protein in directing integration specificity to be explored (13). Deletion of the PWWP chromatin-binding region of LEDGF reduces HIV-1 replication to the level observed in the absence of this cofactor, but replacing the PWWP domain with other chromatin-binding peptides restores efficient replication (14).…”

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confidence: 99%

Targeting HIV-1 DNA integration by swapping tethers

Craigie

2010

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

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LEDGF/p75 Proteins with Alternative Chromatin Tethers Are Functional HIV-1 Cofactors

Cited by 61 publications

References 70 publications

Role of the PWWP Domain of Lens Epithelium-derived Growth Factor (LEDGF)/p75 Cofactor in Lentiviral Integration Targeting

Role of the PWWP Domain of Lens Epithelium-derived Growth Factor (LEDGF)/p75 Cofactor in Lentiviral Integration Targeting

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

Targeting HIV-1 DNA integration by swapping tethers

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