Article: Mutational analysis of the "turn" of helix clamp motif of HIV-1 reverse transcriptase

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TitleMutational analysis of the "turn" of helix clamp motif of HIV-1 reverse transcriptase
AuthorsWang, YX2
Zhang, HJ1
Xu, J2
Zheng, BJ1
Wen, YM2
Issue Date2008
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
CitationBiochemical And Biophysical Research Communications, 2008, v. 377 n. 3, p. 915-920 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.bbrc.2008.10.106
AbstractHelix clamp motifs of polymerases possessing the helix-turn-helix secondary structure are crucial for their polymerase activity by binding to the nucleic acid template/primer via the alpha helices. To study the functions of turn in helix clamp motif of human immunodeficiency virus (HIV)-1 RT, clones with turn mutants at rt271-274 of HIV-1 RT were generated and studied by one cycle infection assay. Mutants rtY271A and rtI274A almost lost their replication competency, while mutants rtA272P, rtA272S, and rtG273A retained comparable replication competency relative to wild type pseudotyped HIV-1. To study the mechanisms involved, RT proteins from rt271 to rt274 mutants were expressed and assayed for their RNA dependent DNA polymerase activity, DNA binding activity and processivity. Discordance between RT activity and viral replication efficiency of some turn mutants was observed, indicating that aside from RT, other steps in HIV replication could be affected by substitutions at the turn of helix clamp motif. © 2008 Elsevier Inc. All rights reserved.
ISSN0006-291X
2011 Impact Factor: 2.484
2011 SCImago Journal Rankings: 0.287
DOIhttp://dx.doi.org/10.1016/j.bbrc.2008.10.106
ISI Accession Number IDWOS:000261458900034
Funding AgencyGrant Number
Shanghai Educational Development FoundationKBF101038
NSFC30800048
Sino-Germen Grant GZ230202/3
Shanghai Municipal Government Basic Research05JC14008
AIDS Trust Fund of Hong Kong SAR Government
Funding Information:

Jurkat Clone E6-1, pHEF-VSVG, and pNL4-3-deltaE-EGFP were kindly provided by the AIDS Research and Reference Reagent Program. This study was supported in part by Grants from 973 (G1999054105), Shanghai Educational Development Foundation (KBF101038), NSFC (30800048), Sino-Germen Grant GZ230 (202/3), Shanghai Municipal Government Basic Research (05JC14008), and the AIDS Trust Fund of Hong Kong SAR Government.

ReferencesReferences in Scopus
DC Field
Value
dc.contributor.authorWang, YX
dc.contributor.authorZhang, HJ
dc.contributor.authorXu, J
dc.contributor.authorZheng, BJ
dc.contributor.authorWen, YM
dc.date.accessioned2012-08-08T08:51:01Z
dc.date.available2012-08-08T08:51:01Z
dc.date.issued2008
dc.description.abstractHelix clamp motifs of polymerases possessing the helix-turn-helix secondary structure are crucial for their polymerase activity by binding to the nucleic acid template/primer via the alpha helices. To study the functions of turn in helix clamp motif of human immunodeficiency virus (HIV)-1 RT, clones with turn mutants at rt271-274 of HIV-1 RT were generated and studied by one cycle infection assay. Mutants rtY271A and rtI274A almost lost their replication competency, while mutants rtA272P, rtA272S, and rtG273A retained comparable replication competency relative to wild type pseudotyped HIV-1. To study the mechanisms involved, RT proteins from rt271 to rt274 mutants were expressed and assayed for their RNA dependent DNA polymerase activity, DNA binding activity and processivity. Discordance between RT activity and viral replication efficiency of some turn mutants was observed, indicating that aside from RT, other steps in HIV replication could be affected by substitutions at the turn of helix clamp motif. © 2008 Elsevier Inc. All rights reserved.
dc.description.natureLink_to_subscribed_fulltext
dc.identifier.citationBiochemical And Biophysical Research Communications, 2008, v. 377 n. 3, p. 915-920 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.bbrc.2008.10.106
dc.identifier.doihttp://dx.doi.org/10.1016/j.bbrc.2008.10.106
dc.identifier.epage920
dc.identifier.hkuros156479
dc.identifier.hkuros168239
dc.identifier.isiWOS:000261458900034
Funding AgencyGrant Number
Shanghai Educational Development FoundationKBF101038
NSFC30800048
Sino-Germen Grant GZ230202/3
Shanghai Municipal Government Basic Research05JC14008
AIDS Trust Fund of Hong Kong SAR Government
Funding Information:

Jurkat Clone E6-1, pHEF-VSVG, and pNL4-3-deltaE-EGFP were kindly provided by the AIDS Research and Reference Reagent Program. This study was supported in part by Grants from 973 (G1999054105), Shanghai Educational Development Foundation (KBF101038), NSFC (30800048), Sino-Germen Grant GZ230 (202/3), Shanghai Municipal Government Basic Research (05JC14008), and the AIDS Trust Fund of Hong Kong SAR Government.

dc.identifier.issn0006-291X
2011 Impact Factor: 2.484
2011 SCImago Journal Rankings: 0.287
dc.identifier.issue3
dc.identifier.pmid18976635
dc.identifier.scopuseid_2-s2.0-56049097627
dc.identifier.spage915
dc.identifier.urihttp://hdl.handle.net/10722/157533
dc.identifier.volume377
dc.languageeng
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
dc.publisher.placeUnited States
dc.relation.ispartofBiochemical and Biophysical Research Communications
dc.relation.referencesReferences in Scopus
dc.subject.meshAmino Acid Motifs - Genetics
dc.subject.meshAmino Acid Substitution
dc.subject.meshCell Line
dc.subject.meshDna - Metabolism
dc.subject.meshHiv Reverse Transcriptase - Genetics - Metabolism
dc.subject.meshHiv-1 - Enzymology - Genetics - Physiology
dc.subject.meshHumans
dc.subject.meshMutation
dc.subject.meshProtein Structure, Secondary
dc.subject.meshVirus Replication - Genetics
dc.titleMutational analysis of the "turn" of helix clamp motif of HIV-1 reverse transcriptase
dc.typeArticle
Author Affiliations
  1. The University of Hong Kong
  2. Fudan University Shanghai Medical College