Eukaryotic elongation factor 1 complex subunits are critical HIV-1 reverse transcription cofactors

Warren, Kylie, Wei, Ting, Li, Dongsheng, Qin, Fangyun, Warrilow, David, Lin, Min-Hsuan, Sivakumaran, Haran, Apolloni, Ann, Abbott, Catherine M., Jones, Alun, Anderson, Jenny L. and Harrich, David (2012) Eukaryotic elongation factor 1 complex subunits are critical HIV-1 reverse transcription cofactors. Proceedings of the National Academy of Sciences of USA, 109 24: 9587-9592. doi:10.1073/pnas.1204673109

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Author Warren, Kylie
Wei, Ting
Li, Dongsheng
Qin, Fangyun
Warrilow, David
Lin, Min-Hsuan
Sivakumaran, Haran
Apolloni, Ann
Abbott, Catherine M.
Jones, Alun
Anderson, Jenny L.
Harrich, David
Title Eukaryotic elongation factor 1 complex subunits are critical HIV-1 reverse transcription cofactors
Journal name Proceedings of the National Academy of Sciences of USA   Check publisher's open access policy
ISSN 0027-8424
1091-6490
Publication date 2012-06-12
Sub-type Article (original research)
DOI 10.1073/pnas.1204673109
Open Access Status File (Publisher version)
Volume 109
Issue 24
Start page 9587
End page 9592
Total pages 6
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Collection year 2013
Language eng
Abstract Cellular proteins have been implicated as important for HIV-1 reverse transcription, but whether any are reverse transcription complex (RTC) cofactors or affect reverse transcription indirectly is unclear. Here we used protein fractionation combined with an endogenous reverse transcription assay to identify cellular proteins that stimulated late steps of reverse transcription in vitro. We identified 25 cellular proteins in an active protein fraction, and here we show that the eEF1A and eEF1G subunits of eukaryotic elongation factor 1 (eEF1) are important components of the HIV-1 RTC. eEF1A and eEF1G were identified in fractionated human T-cell lysates as reverse transcription cofactors, as their removal ablated the ability of active protein fractions to stimulate late reverse transcription in vitro. We observed that the p51 subunit of reverse transcriptase and integrase, two subunits of the RTC, coimmunoprecipitated with eEF1A and eEF1G. Moreover eEF1A and eEF1G associated with purified RTCs and colocalized with reverse transcriptase following infection of cells. Reverse transcription in cells was sharply down-regulated when eEF1A or eEF1G levels were reduced by siRNA treatment as a result of reduced levels of RTCs in treated cells. The combined evidence indicates that these eEF1 subunits are critical RTC stability cofactors required for efficient completion of reverse transcription. The identification of eEF1 subunits as unique RTC components provides a basis for further investigations of reverse transcription and trafficking of the RTC to the nucleus.
Keyword Purification
Mass spectrometry
Cellular factor
siRNA knock down
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Chemistry and Molecular Biosciences
Institute for Molecular Bioscience - Publications
 
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Created: Wed, 04 Jul 2012, 11:04:23 EST by Lucy O'Brien on behalf of School of Chemistry & Molecular Biosciences