Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.

Val75 of HIV-1 reverse transcriptase (RT) plays a role in positioning the template nucleotide +1, during the formation of the ternary complex. Mutations such as V75M and V75A emerge in patients infected with HIV-1 group M-subtype B and group O variants, after failing treatment with stavudine (d4T) a...

ver descrição completa

Detalhes bibliográficos
Autores: Matamoros Grande, Tania, Nevot, María, Martínez, Miguel Ángel, Menéndez-Arias, Luis
Formato: artículo
Fecha de publicación:2009
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/24852
Acesso em linha:http://hdl.handle.net/10261/24852
Access Level:acceso abierto
Palavra-chave:HIV-1 reverse transcriptase (RT)
thymidine analogue resistance mutations (TAMs)
id ES_6e0b463028fa9f72bc729a3e41126a6a
oai_identifier_str oai:digital.csic.es:10261/24852
network_acronym_str ES
network_name_str España
repository_id_str
spelling Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.Matamoros Grande, TaniaNevot, MaríaMartínez, Miguel ÁngelMenéndez-Arias, LuisHIV-1 reverse transcriptase (RT)thymidine analogue resistance mutations (TAMs)Val75 of HIV-1 reverse transcriptase (RT) plays a role in positioning the template nucleotide +1, during the formation of the ternary complex. Mutations such as V75M and V75A emerge in patients infected with HIV-1 group M-subtype B and group O variants, after failing treatment with stavudine (d4T) and other nucleoside RT inhibitors. V75I is an accessory mutation of the Q151M multi-drug resistance complex of HIV-1 RT, and is rarely associated with thymidine analogue resistance mutations (TAMs). In vitro, it confers resistance to acyclovir. TAMs confer resistance to zidovudine (AZT) and d4T by increasing the rate of ATPmediated excision of the terminal nucleotide monophosphate (primer unblocking). In a wild-type HIV-1 group O RT sequence context, V75A and V75M conferred increased excision activity on d4T-terminated primers, in the presence of pyrophosphate (PPi). In contrast, V75I decreased the PPi-mediated unblocking efficiency on AZT and d4T-terminated primers, in different sequence contexts (i.e. wild-type group M-subtype B or group O RTs). Interestingly, in the sequence context of an excision-proficient RT (i.e., M41L/A62V/T69SSS/K70R/T215Y), the introduction of V75I led to a significant decrease of its ATP-dependent excision activity on AZT-, d4T-, and acyclovirterminated primers. The excision rate of d4T-monophosphate in the presence of ATP 3.2 mM was about 10 times higher for M41L/A62V/T69SSS/K70R/T215Y than for the mutant M41L/A62V/T69SSS/K70R/- V75I/T215Y RT. The antagonistic effect of V75I with TAMs was further demonstrated in phenotypic assays. Recombinant HIV-1 containing the M41L/A62V/T69SSS/- K70R/V75I/T215Y RT showed 18.3- and 1.5-fold increased susceptibility to AZT and d4T, respectively, in comparison with virus containing the M41L/A62V/T69SSS/- K70R/T215Y RT.This work was supported by Spanish Ministry of Science and Innovation Grant BIO2007/60319, Fundacio´n para la Investigacio´n y Prevencio´n del SIDA en Espan˜ a (FIPSE) Grant 36771/08, the Fondo de Investigacio´n Sanitaria (through “Red Tema´tica de Investigacio´n Cooperativa en SIDA” Grant RD06/0006), and an institutional grant from the Fundacio´n Ramo´n Areces. This work was also supported by Spanish Ministry of Science and Innovation Grants BFU2006/01066 and PI07/0098 (to M. A. M.)Peer reviewedAmerican Society for Biochemistry and Molecular BiologyMinisterio de Ciencia e Innovación (España)Fundación para la Investigación y la Prevención del Sida en EspañaInstituto de Salud Carlos IIIFundación Ramón Areces201020102009info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501154379 bytesapplication/pdfhttp://hdl.handle.net/10261/24852reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1074/jbc.M109.038885info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/248522026-05-22T06:33:51Z
dc.title.none.fl_str_mv Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
title Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
spellingShingle Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
Matamoros Grande, Tania
HIV-1 reverse transcriptase (RT)
thymidine analogue resistance mutations (TAMs)
title_short Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
title_full Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
title_fullStr Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
title_full_unstemmed Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
title_sort Thymidine Analogue Resistance Suppression By V75i Of Hiv-1 Reverse Transcriptase: Effects Of Substituting Valine 75 On Stavudine Excision And Discrimination.
dc.creator.none.fl_str_mv Matamoros Grande, Tania
Nevot, María
Martínez, Miguel Ángel
Menéndez-Arias, Luis
author Matamoros Grande, Tania
author_facet Matamoros Grande, Tania
Nevot, María
Martínez, Miguel Ángel
Menéndez-Arias, Luis
author_role author
author2 Nevot, María
Martínez, Miguel Ángel
Menéndez-Arias, Luis
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Fundación para la Investigación y la Prevención del Sida en España
Instituto de Salud Carlos III
Fundación Ramón Areces
dc.subject.none.fl_str_mv HIV-1 reverse transcriptase (RT)
thymidine analogue resistance mutations (TAMs)
topic HIV-1 reverse transcriptase (RT)
thymidine analogue resistance mutations (TAMs)
description Val75 of HIV-1 reverse transcriptase (RT) plays a role in positioning the template nucleotide +1, during the formation of the ternary complex. Mutations such as V75M and V75A emerge in patients infected with HIV-1 group M-subtype B and group O variants, after failing treatment with stavudine (d4T) and other nucleoside RT inhibitors. V75I is an accessory mutation of the Q151M multi-drug resistance complex of HIV-1 RT, and is rarely associated with thymidine analogue resistance mutations (TAMs). In vitro, it confers resistance to acyclovir. TAMs confer resistance to zidovudine (AZT) and d4T by increasing the rate of ATPmediated excision of the terminal nucleotide monophosphate (primer unblocking). In a wild-type HIV-1 group O RT sequence context, V75A and V75M conferred increased excision activity on d4T-terminated primers, in the presence of pyrophosphate (PPi). In contrast, V75I decreased the PPi-mediated unblocking efficiency on AZT and d4T-terminated primers, in different sequence contexts (i.e. wild-type group M-subtype B or group O RTs). Interestingly, in the sequence context of an excision-proficient RT (i.e., M41L/A62V/T69SSS/K70R/T215Y), the introduction of V75I led to a significant decrease of its ATP-dependent excision activity on AZT-, d4T-, and acyclovirterminated primers. The excision rate of d4T-monophosphate in the presence of ATP 3.2 mM was about 10 times higher for M41L/A62V/T69SSS/K70R/T215Y than for the mutant M41L/A62V/T69SSS/K70R/- V75I/T215Y RT. The antagonistic effect of V75I with TAMs was further demonstrated in phenotypic assays. Recombinant HIV-1 containing the M41L/A62V/T69SSS/- K70R/V75I/T215Y RT showed 18.3- and 1.5-fold increased susceptibility to AZT and d4T, respectively, in comparison with virus containing the M41L/A62V/T69SSS/- K70R/T215Y RT.
publishDate 2009
dc.date.none.fl_str_mv 2009
2010
2010
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/24852
url http://hdl.handle.net/10261/24852
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1074/jbc.M109.038885
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 154379 bytes
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869410396487548928
score 15,812429

Registos relacionados