HIV-1 reverse transcriptase connection subdomain mutations reduce template RNA degradation and enhance AZT excision
We previously proposed that mutations in the connection subdomain (cn) of HIV-1 reverse transcriptase increase AZT resistance by altering the balance between nucleotide excision and template RNA degradation. To test the predictions of this model, we analyzed the effects of previously identified cn m...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2008-08, Vol.105 (31), p.10943-10948 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Delviks-Frankenberry, Krista A Nikolenko, Galina N Boyer, Paul L Hughes, Stephen H Coffin, John M Jere, Abhay Pathak, Vinay K |
| description | We previously proposed that mutations in the connection subdomain (cn) of HIV-1 reverse transcriptase increase AZT resistance by altering the balance between nucleotide excision and template RNA degradation. To test the predictions of this model, we analyzed the effects of previously identified cn mutations in combination with thymidine analog mutations (D67N, K70R, T215Y, and K219Q) on in vitro RNase H activity and AZT monophosphate (AZTMP) excision. We found that cn mutations G335C/D, N348I, A360I/V, V365I, and A376S decreased primary and secondary RNase H cleavages. The patient-derived cns increased ATP- and PPi-mediated AZTMP excision on an RNA template compared with a DNA template. One of 5 cns caused an increase in ATP-mediated AZTMP excision on a DNA template, whereas three cns showed a higher ratio of ATP- to PPi-mediated excision, indicating that some cn mutations also affect excision on a DNA substrate. Overall, the results strongly support the model that cn mutations increase AZT resistance by reducing template RNA degradation, thereby providing additional time for RT to excise AZTMP. |
| doi_str_mv | 10.1073/pnas.0804660105 |
| format | Article |
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To test the predictions of this model, we analyzed the effects of previously identified cn mutations in combination with thymidine analog mutations (D67N, K70R, T215Y, and K219Q) on in vitro RNase H activity and AZT monophosphate (AZTMP) excision. We found that cn mutations G335C/D, N348I, A360I/V, V365I, and A376S decreased primary and secondary RNase H cleavages. The patient-derived cns increased ATP- and PPi-mediated AZTMP excision on an RNA template compared with a DNA template. One of 5 cns caused an increase in ATP-mediated AZTMP excision on a DNA template, whereas three cns showed a higher ratio of ATP- to PPi-mediated excision, indicating that some cn mutations also affect excision on a DNA substrate. Overall, the results strongly support the model that cn mutations increase AZT resistance by reducing template RNA degradation, thereby providing additional time for RT to excise AZTMP.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.0804660105</identifier><identifier>PMID: 18667707</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Biological Sciences ; Cell Line ; Cloning, Molecular ; Deoxyribonucleic acid ; DNA ; DNA Primers - genetics ; Drug resistance ; Drug Resistance, Viral - genetics ; Genetic mutation ; Genetic vectors ; HIV ; HIV 1 ; HIV Reverse Transcriptase - genetics ; Human immunodeficiency virus ; Human immunodeficiency virus 1 ; Humans ; Hybridity ; Impact analysis ; Models, Biological ; Mutagenesis ; Mutation ; Mutation - genetics ; Polymerization ; Ribonucleic acid ; RNA ; RNA - metabolism ; RNA stability ; Viruses ; Zidovudine - metabolism</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2008-08, Vol.105 (31), p.10943-10948</ispartof><rights>Copyright 2008 The National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Aug 5, 2008</rights><rights>2008 by The National Academy of Sciences of the USA</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c620t-8d0bd668c3662af36d386d755a797d34b456d6cf6c569289388a5626f9c6c23e3</citedby><cites>FETCH-LOGICAL-c620t-8d0bd668c3662af36d386d755a797d34b456d6cf6c569289388a5626f9c6c23e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/105/31.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/25463266$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/25463266$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18667707$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Delviks-Frankenberry, Krista A</creatorcontrib><creatorcontrib>Nikolenko, Galina N</creatorcontrib><creatorcontrib>Boyer, Paul L</creatorcontrib><creatorcontrib>Hughes, Stephen H</creatorcontrib><creatorcontrib>Coffin, John M</creatorcontrib><creatorcontrib>Jere, Abhay</creatorcontrib><creatorcontrib>Pathak, Vinay K</creatorcontrib><title>HIV-1 reverse transcriptase connection subdomain mutations reduce template RNA degradation and enhance AZT excision</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>We previously proposed that mutations in the connection subdomain (cn) of HIV-1 reverse transcriptase increase AZT resistance by altering the balance between nucleotide excision and template RNA degradation. To test the predictions of this model, we analyzed the effects of previously identified cn mutations in combination with thymidine analog mutations (D67N, K70R, T215Y, and K219Q) on in vitro RNase H activity and AZT monophosphate (AZTMP) excision. We found that cn mutations G335C/D, N348I, A360I/V, V365I, and A376S decreased primary and secondary RNase H cleavages. The patient-derived cns increased ATP- and PPi-mediated AZTMP excision on an RNA template compared with a DNA template. One of 5 cns caused an increase in ATP-mediated AZTMP excision on a DNA template, whereas three cns showed a higher ratio of ATP- to PPi-mediated excision, indicating that some cn mutations also affect excision on a DNA substrate. Overall, the results strongly support the model that cn mutations increase AZT resistance by reducing template RNA degradation, thereby providing additional time for RT to excise AZTMP.</description><subject>Biological Sciences</subject><subject>Cell Line</subject><subject>Cloning, Molecular</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Primers - genetics</subject><subject>Drug resistance</subject><subject>Drug Resistance, Viral - genetics</subject><subject>Genetic mutation</subject><subject>Genetic vectors</subject><subject>HIV</subject><subject>HIV 1</subject><subject>HIV Reverse Transcriptase - genetics</subject><subject>Human immunodeficiency virus</subject><subject>Human immunodeficiency virus 1</subject><subject>Humans</subject><subject>Hybridity</subject><subject>Impact analysis</subject><subject>Models, Biological</subject><subject>Mutagenesis</subject><subject>Mutation</subject><subject>Mutation - genetics</subject><subject>Polymerization</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA - metabolism</subject><subject>RNA stability</subject><subject>Viruses</subject><subject>Zidovudine - metabolism</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFks1v1DAQxSMEokvhzAmIOCBxSDv-mjgXpFUFtFIFErQcuFhe29lmldiLnVTlv8dhV13gwmlkz-89-em5KJ4TOCFQs9Ot1-kEJHBEICAeFAsCDamQN_CwWADQupKc8qPiSUobAGiEhMfFEZGIdQ31okjnF98qUkZ362Jy5Ri1TyZ221HnkwneOzN2wZdpWtkw6M6XwzTq-SplkZ1M1rhh2-vRlV8-LUvr1lHb30CpvS2dv9E-Q8vvV6W7M13Ki6fFo1b3yT3bz-Pi-sP7q7Pz6vLzx4uz5WVlkMJYSQsriygNQ6S6ZWiZRFsLoeumtoyvuECLpkUjsKGyYVJqgRTbxqChzLHj4t3OdzutBmeN8zler7axG3T8qYLu1N8b392odbhVlDeES5kN3uwNYvgxuTSqoUvG9b32LkxJkQaBEQEZfP0PuAlT9DmcokCYFDWZ3U53kIkhpeja-5cQUHObam5THdrMipd_Bjjw-_oyUO6BWXmwE4qRPBrOMvL2P4hqp74f3d2Y2Rc7dpPGEO9hKjgyipj3r3b7Vgel17FL6vrrHDB_LEq5IOwXNLnH1A</recordid><startdate>20080805</startdate><enddate>20080805</enddate><creator>Delviks-Frankenberry, Krista A</creator><creator>Nikolenko, Galina N</creator><creator>Boyer, Paul L</creator><creator>Hughes, Stephen H</creator><creator>Coffin, John M</creator><creator>Jere, Abhay</creator><creator>Pathak, Vinay K</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20080805</creationdate><title>HIV-1 reverse transcriptase connection subdomain mutations reduce template RNA degradation and enhance AZT excision</title><author>Delviks-Frankenberry, Krista A ; 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To test the predictions of this model, we analyzed the effects of previously identified cn mutations in combination with thymidine analog mutations (D67N, K70R, T215Y, and K219Q) on in vitro RNase H activity and AZT monophosphate (AZTMP) excision. We found that cn mutations G335C/D, N348I, A360I/V, V365I, and A376S decreased primary and secondary RNase H cleavages. The patient-derived cns increased ATP- and PPi-mediated AZTMP excision on an RNA template compared with a DNA template. One of 5 cns caused an increase in ATP-mediated AZTMP excision on a DNA template, whereas three cns showed a higher ratio of ATP- to PPi-mediated excision, indicating that some cn mutations also affect excision on a DNA substrate. Overall, the results strongly support the model that cn mutations increase AZT resistance by reducing template RNA degradation, thereby providing additional time for RT to excise AZTMP.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>18667707</pmid><doi>10.1073/pnas.0804660105</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Biological Sciences Cell Line Cloning, Molecular Deoxyribonucleic acid DNA DNA Primers - genetics Drug resistance Drug Resistance, Viral - genetics Genetic mutation Genetic vectors HIV HIV 1 HIV Reverse Transcriptase - genetics Human immunodeficiency virus Human immunodeficiency virus 1 Humans Hybridity Impact analysis Models, Biological Mutagenesis Mutation Mutation - genetics Polymerization Ribonucleic acid RNA RNA - metabolism RNA stability Viruses Zidovudine - metabolism |
| title | HIV-1 reverse transcriptase connection subdomain mutations reduce template RNA degradation and enhance AZT excision |
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