Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.

Recent work suggests a role for multiple host factors in facilitating HIV-1 reverse transcription. Previously, we identified a cellular activity which increases the efficiency of HIV-1 reverse transcription in vitro. Here, we describe aspects of the activity which shed light on its function. The cel...

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Autores principales:	David Warrilow, Kylie Warren, David Harrich
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9bf2403804354526be33485bf01b749e
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spelling	oai:doaj.org-article:9bf2403804354526be33485bf01b749e2021-11-18T07:03:38ZStrand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.1932-620310.1371/journal.pone.0013229https://doaj.org/article/9bf2403804354526be33485bf01b749e2010-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20949087/?tool=EBIhttps://doaj.org/toc/1932-6203Recent work suggests a role for multiple host factors in facilitating HIV-1 reverse transcription. Previously, we identified a cellular activity which increases the efficiency of HIV-1 reverse transcription in vitro. Here, we describe aspects of the activity which shed light on its function. The cellular factor did not affect synthesis of strong-stop DNA but did improve downstream DNA synthesis. The stimulatory activity was isolated by gel filtration in a single fraction of the exclusion volume. Velocity-gradient purified HIV-1, which was free of detectable RNase activity, showed poor reverse transcription efficiency but was strongly stimulated by partially purified cell proteins. Hence, the cell factor(s) did not inactivate an RNase activity that might degrade the viral genomic RNA and block completion of reverse transcription. Instead, the cell factor(s) enhanced first strand transfer and synthesis of late reverse transcription suggesting it stabilized the reverse transcription complex. The factor did not affect lysis of HIV-1 by Triton X-100 in the endogenous reverse transcription (ERT) system, and ERT reactions with HIV-1 containing capsid mutations, which varied the biochemical stability of viral core structures and impeded reverse transcription in cells, showed no difference in the ability to be stimulated by the cell factor(s) suggesting a lack of involvement of the capsid in the in vitro assay. In addition, reverse transcription products were found to be resistant to exogenous DNase I activity when the active fraction was present in the ERT assay. These results indicate that the cell factor(s) may improve reverse transcription by facilitating DNA strand transfer and DNA synthesis. It also had a protective function for the reverse transcription products, but it is unclear if this is related to improved DNA synthesis.David WarrilowKylie WarrenDavid HarrichPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 10, p e13229 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q David Warrilow Kylie Warren David Harrich Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
description	Recent work suggests a role for multiple host factors in facilitating HIV-1 reverse transcription. Previously, we identified a cellular activity which increases the efficiency of HIV-1 reverse transcription in vitro. Here, we describe aspects of the activity which shed light on its function. The cellular factor did not affect synthesis of strong-stop DNA but did improve downstream DNA synthesis. The stimulatory activity was isolated by gel filtration in a single fraction of the exclusion volume. Velocity-gradient purified HIV-1, which was free of detectable RNase activity, showed poor reverse transcription efficiency but was strongly stimulated by partially purified cell proteins. Hence, the cell factor(s) did not inactivate an RNase activity that might degrade the viral genomic RNA and block completion of reverse transcription. Instead, the cell factor(s) enhanced first strand transfer and synthesis of late reverse transcription suggesting it stabilized the reverse transcription complex. The factor did not affect lysis of HIV-1 by Triton X-100 in the endogenous reverse transcription (ERT) system, and ERT reactions with HIV-1 containing capsid mutations, which varied the biochemical stability of viral core structures and impeded reverse transcription in cells, showed no difference in the ability to be stimulated by the cell factor(s) suggesting a lack of involvement of the capsid in the in vitro assay. In addition, reverse transcription products were found to be resistant to exogenous DNase I activity when the active fraction was present in the ERT assay. These results indicate that the cell factor(s) may improve reverse transcription by facilitating DNA strand transfer and DNA synthesis. It also had a protective function for the reverse transcription products, but it is unclear if this is related to improved DNA synthesis.
format	article
author	David Warrilow Kylie Warren David Harrich
author_facet	David Warrilow Kylie Warren David Harrich
author_sort	David Warrilow
title	Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
title_short	Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
title_full	Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
title_fullStr	Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
title_full_unstemmed	Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.
title_sort	strand transfer and elongation of hiv-1 reverse transcription is facilitated by cell factors in vitro.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/9bf2403804354526be33485bf01b749e
work_keys_str_mv	AT davidwarrilow strandtransferandelongationofhiv1reversetranscriptionisfacilitatedbycellfactorsinvitro AT kyliewarren strandtransferandelongationofhiv1reversetranscriptionisfacilitatedbycellfactorsinvitro AT davidharrich strandtransferandelongationofhiv1reversetranscriptionisfacilitatedbycellfactorsinvitro
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Strand transfer and elongation of HIV-1 reverse transcription is facilitated by cell factors in vitro.

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