Structure-based mutagenesis of the integrase-LEDGF/p75 interface uncouples a strict correlation between in vitro protein binding and HIV-1 fitness
Abstract LEDGF/p75 binding-defective IN mutant viruses were previously characterized as replication-defective, yet RNAi did not reveal an essential role for the host factor in HIV-1 replication. Correlative analyses of protein binding and viral fitness were expanded here by targeting 12 residues at...
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| Veröffentlicht in: | Virology (New York, N.Y.) N.Y.), 2007-01, Vol.357 (1), p.79-90 |
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| creator | Rahman, Shaila Lu, Richard Vandegraaff, Nick Cherepanov, Peter Engelman, Alan |
| description | Abstract LEDGF/p75 binding-defective IN mutant viruses were previously characterized as replication-defective, yet RNAi did not reveal an essential role for the host factor in HIV-1 replication. Correlative analyses of protein binding and viral fitness were expanded here by targeting 12 residues at the IN-LEDGF/p75 binding interface. Whereas many of the resultant viruses were defective, the majority of the INs displayed wild-type in vitro integration activities. Though an overall trend of parallel loss of LEDGF/p75 binding and HIV-1 infectivity was observed, a strict correlation was not. His-tagged INA128Q , derived from a phenotypically wild-type virus, failed to pull-down LEDGF/p75, but INA128Q was effectively recovered in a reciprocal GST pull-down assay. Under these conditions, INH171A , also derived from a phenotypically wild-type virus, interacted less efficiently than a previously described interaction-defective mutant, INQ168A . Thus, the relative affinity of the in vitro IN-LEDGF/p75 interaction is not a universal predictor of IN mutant viral fitness. |
| doi_str_mv | 10.1016/j.virol.2006.08.011 |
| format | Article |
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Correlative analyses of protein binding and viral fitness were expanded here by targeting 12 residues at the IN-LEDGF/p75 binding interface. Whereas many of the resultant viruses were defective, the majority of the INs displayed wild-type in vitro integration activities. Though an overall trend of parallel loss of LEDGF/p75 binding and HIV-1 infectivity was observed, a strict correlation was not. His-tagged INA128Q , derived from a phenotypically wild-type virus, failed to pull-down LEDGF/p75, but INA128Q was effectively recovered in a reciprocal GST pull-down assay. Under these conditions, INH171A , also derived from a phenotypically wild-type virus, interacted less efficiently than a previously described interaction-defective mutant, INQ168A . Thus, the relative affinity of the in vitro IN-LEDGF/p75 interaction is not a universal predictor of IN mutant viral fitness.</description><identifier>ISSN: 0042-6822</identifier><identifier>EISSN: 1096-0341</identifier><identifier>DOI: 10.1016/j.virol.2006.08.011</identifier><identifier>PMID: 16959283</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>AIDS ; Cell Line ; HeLa Cells ; HIV Infections - virology ; HIV Integrase - chemistry ; HIV Integrase - genetics ; HIV Integrase - metabolism ; HIV-1 ; HIV-1 - metabolism ; HIV-1 - physiology ; Human immunodeficiency virus 1 ; Humans ; Infectious Disease ; Integrase ; Integration ; Intercellular Signaling Peptides and Proteins - metabolism ; Jurkat Cells ; LEDGF/p75 ; Models, Molecular ; Mutagenesis ; Protein Binding ; Protein interaction ; Virus Integration</subject><ispartof>Virology (New York, N.Y.), 2007-01, Vol.357 (1), p.79-90</ispartof><rights>Elsevier Inc.</rights><rights>2006 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c554t-d662797b701c7078ecba15122f51dd0f01019a80742b6598c19bbcb1bfd0a4ad3</citedby><cites>FETCH-LOGICAL-c554t-d662797b701c7078ecba15122f51dd0f01019a80742b6598c19bbcb1bfd0a4ad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0042682206005575$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16959283$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rahman, Shaila</creatorcontrib><creatorcontrib>Lu, Richard</creatorcontrib><creatorcontrib>Vandegraaff, Nick</creatorcontrib><creatorcontrib>Cherepanov, Peter</creatorcontrib><creatorcontrib>Engelman, Alan</creatorcontrib><title>Structure-based mutagenesis of the integrase-LEDGF/p75 interface uncouples a strict correlation between in vitro protein binding and HIV-1 fitness</title><title>Virology (New York, N.Y.)</title><addtitle>Virology</addtitle><description>Abstract LEDGF/p75 binding-defective IN mutant viruses were previously characterized as replication-defective, yet RNAi did not reveal an essential role for the host factor in HIV-1 replication. Correlative analyses of protein binding and viral fitness were expanded here by targeting 12 residues at the IN-LEDGF/p75 binding interface. Whereas many of the resultant viruses were defective, the majority of the INs displayed wild-type in vitro integration activities. Though an overall trend of parallel loss of LEDGF/p75 binding and HIV-1 infectivity was observed, a strict correlation was not. His-tagged INA128Q , derived from a phenotypically wild-type virus, failed to pull-down LEDGF/p75, but INA128Q was effectively recovered in a reciprocal GST pull-down assay. Under these conditions, INH171A , also derived from a phenotypically wild-type virus, interacted less efficiently than a previously described interaction-defective mutant, INQ168A . Thus, the relative affinity of the in vitro IN-LEDGF/p75 interaction is not a universal predictor of IN mutant viral fitness.</description><subject>AIDS</subject><subject>Cell Line</subject><subject>HeLa Cells</subject><subject>HIV Infections - virology</subject><subject>HIV Integrase - chemistry</subject><subject>HIV Integrase - genetics</subject><subject>HIV Integrase - metabolism</subject><subject>HIV-1</subject><subject>HIV-1 - metabolism</subject><subject>HIV-1 - physiology</subject><subject>Human immunodeficiency virus 1</subject><subject>Humans</subject><subject>Infectious Disease</subject><subject>Integrase</subject><subject>Integration</subject><subject>Intercellular Signaling Peptides and Proteins - metabolism</subject><subject>Jurkat Cells</subject><subject>LEDGF/p75</subject><subject>Models, Molecular</subject><subject>Mutagenesis</subject><subject>Protein Binding</subject><subject>Protein interaction</subject><subject>Virus Integration</subject><issn>0042-6822</issn><issn>1096-0341</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUsuOEzEQtBCIDQtfgIR84jazbU88jwNIaN9SJA4LXC0_eoLDxA62J2h_gy_G2URC4sKp1e2q7nZVE_KWQc2AtRebeu9imGoO0NbQ18DYM7JgMLQVNEv2nCwAlrxqe87PyKuUNlDyroOX5Iy1gxh43yzI74ccZ5PniJVWCS3dzlmt0WNyiYaR5u9Inc-4juW1Wl1f3d5c7DrxVIujMkhnb8K8mzBRRVOOzmRqQow4qeyCpxrzL0RfCHTvcgx0F0PGkmnnrfNrqryld_ffKkZHl8vc9Jq8GNWU8M0pnpOvN9dfLu-q1efb-8tPq8oIscyVbVveDZ3ugJkOuh6NVkwwzkfBrIURikiD6qFbct2KoTds0NpopkcLaqlsc07eH_uWjX7OmLLcumRwmpTHMCfJBsEF500BNkegiSGliKPcRbdV8VEykAcr5EY-WSEPVkjoZbGisN6d2s96i_Yv56R9AXw4ArB8cu8wymQceoPWRTRZ2uD-M-DjP3wzOe-Mmn7gI6ZNmKMv-kkmE5cgHw7XcDgGaAGE6ETzB81Lsj4</recordid><startdate>20070105</startdate><enddate>20070105</enddate><creator>Rahman, Shaila</creator><creator>Lu, Richard</creator><creator>Vandegraaff, Nick</creator><creator>Cherepanov, Peter</creator><creator>Engelman, Alan</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20070105</creationdate><title>Structure-based mutagenesis of the integrase-LEDGF/p75 interface uncouples a strict correlation between in vitro protein binding and HIV-1 fitness</title><author>Rahman, Shaila ; Lu, Richard ; Vandegraaff, Nick ; Cherepanov, Peter ; Engelman, Alan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c554t-d662797b701c7078ecba15122f51dd0f01019a80742b6598c19bbcb1bfd0a4ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>AIDS</topic><topic>Cell Line</topic><topic>HeLa Cells</topic><topic>HIV Infections - virology</topic><topic>HIV Integrase - chemistry</topic><topic>HIV Integrase - genetics</topic><topic>HIV Integrase - metabolism</topic><topic>HIV-1</topic><topic>HIV-1 - metabolism</topic><topic>HIV-1 - physiology</topic><topic>Human immunodeficiency virus 1</topic><topic>Humans</topic><topic>Infectious Disease</topic><topic>Integrase</topic><topic>Integration</topic><topic>Intercellular Signaling Peptides and Proteins - metabolism</topic><topic>Jurkat Cells</topic><topic>LEDGF/p75</topic><topic>Models, Molecular</topic><topic>Mutagenesis</topic><topic>Protein Binding</topic><topic>Protein interaction</topic><topic>Virus Integration</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rahman, Shaila</creatorcontrib><creatorcontrib>Lu, Richard</creatorcontrib><creatorcontrib>Vandegraaff, Nick</creatorcontrib><creatorcontrib>Cherepanov, Peter</creatorcontrib><creatorcontrib>Engelman, Alan</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Virology (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rahman, Shaila</au><au>Lu, Richard</au><au>Vandegraaff, Nick</au><au>Cherepanov, Peter</au><au>Engelman, Alan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structure-based mutagenesis of the integrase-LEDGF/p75 interface uncouples a strict correlation between in vitro protein binding and HIV-1 fitness</atitle><jtitle>Virology (New York, N.Y.)</jtitle><addtitle>Virology</addtitle><date>2007-01-05</date><risdate>2007</risdate><volume>357</volume><issue>1</issue><spage>79</spage><epage>90</epage><pages>79-90</pages><issn>0042-6822</issn><eissn>1096-0341</eissn><abstract>Abstract LEDGF/p75 binding-defective IN mutant viruses were previously characterized as replication-defective, yet RNAi did not reveal an essential role for the host factor in HIV-1 replication. Correlative analyses of protein binding and viral fitness were expanded here by targeting 12 residues at the IN-LEDGF/p75 binding interface. Whereas many of the resultant viruses were defective, the majority of the INs displayed wild-type in vitro integration activities. Though an overall trend of parallel loss of LEDGF/p75 binding and HIV-1 infectivity was observed, a strict correlation was not. His-tagged INA128Q , derived from a phenotypically wild-type virus, failed to pull-down LEDGF/p75, but INA128Q was effectively recovered in a reciprocal GST pull-down assay. Under these conditions, INH171A , also derived from a phenotypically wild-type virus, interacted less efficiently than a previously described interaction-defective mutant, INQ168A . 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| subjects | AIDS Cell Line HeLa Cells HIV Infections - virology HIV Integrase - chemistry HIV Integrase - genetics HIV Integrase - metabolism HIV-1 HIV-1 - metabolism HIV-1 - physiology Human immunodeficiency virus 1 Humans Infectious Disease Integrase Integration Intercellular Signaling Peptides and Proteins - metabolism Jurkat Cells LEDGF/p75 Models, Molecular Mutagenesis Protein Binding Protein interaction Virus Integration |
| title | Structure-based mutagenesis of the integrase-LEDGF/p75 interface uncouples a strict correlation between in vitro protein binding and HIV-1 fitness |
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