Fates of retroviral core components during unrestricted and TRIM5-restricted infection.

TRIM5 proteins can restrict retroviral infection soon after delivery of the viral core into the cytoplasm. However, the molecular mechanisms by which TRIM5α inhibits infection have been elusive, in part due to the difficulty of developing and executing biochemical assays that examine this stage of t...

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Autores principales:	Sebla B Kutluay, David Perez-Caballero, Paul D Bieniasz
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/273780abbffe4f918789c7230f890c8b
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spelling	oai:doaj.org-article:273780abbffe4f918789c7230f890c8b2021-11-18T06:05:57ZFates of retroviral core components during unrestricted and TRIM5-restricted infection.1553-73661553-737410.1371/journal.ppat.1003214https://doaj.org/article/273780abbffe4f918789c7230f890c8b2013-03-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23505372/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374TRIM5 proteins can restrict retroviral infection soon after delivery of the viral core into the cytoplasm. However, the molecular mechanisms by which TRIM5α inhibits infection have been elusive, in part due to the difficulty of developing and executing biochemical assays that examine this stage of the retroviral life cycle. Prevailing models suggest that TRIM5α causes premature disassembly of retroviral capsids and/or degradation of capsids by proteasomes, but whether one of these events leads to the other is unclear. Furthermore, how TRIM5α affects the essential components of the viral core, other than capsid, is unknown. To address these questions, we devised a biochemical assay in which the fate of multiple components of retroviral cores during infection can be determined. We utilized cells that can be efficiently infected by VSV-G-pseudotyped retroviruses, and fractionated the cytosolic proteins on linear gradients following synchronized infection. The fates of capsid and integrase proteins, as well as viral genomic RNA and reverse transcription products were then monitored. We found that components of MLV and HIV-1 cores formed a large complex under non-restrictive conditions. In contrast, when MLV infection was restricted by human TRIM5α, the integrase protein and reverse transcription products were lost from infected cells, while capsid and viral RNA were both solubilized. Similarly, when HIV-1 infection was restricted by rhesus TRIM5α or owl monkey TRIMCyp, the integrase protein and reverse transcription products were lost. However, viral RNA was also lost, and high levels of preexisting soluble CA prevented the determination of whether CA was solubilized. Notably, proteasome inhibition blocked all of the aforementioned biochemical consequences of TRIM5α-mediated restriction but had no effect on its antiviral potency. Together, our results show how TRIM5α affects various retroviral core components and indicate that proteasomes are required for TRIM5α-induced core disruption but not for TRIM5α-induced restriction.Sebla B KutluayDavid Perez-CaballeroPaul D BieniaszPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 9, Iss 3, p e1003214 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Sebla B Kutluay David Perez-Caballero Paul D Bieniasz Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
description	TRIM5 proteins can restrict retroviral infection soon after delivery of the viral core into the cytoplasm. However, the molecular mechanisms by which TRIM5α inhibits infection have been elusive, in part due to the difficulty of developing and executing biochemical assays that examine this stage of the retroviral life cycle. Prevailing models suggest that TRIM5α causes premature disassembly of retroviral capsids and/or degradation of capsids by proteasomes, but whether one of these events leads to the other is unclear. Furthermore, how TRIM5α affects the essential components of the viral core, other than capsid, is unknown. To address these questions, we devised a biochemical assay in which the fate of multiple components of retroviral cores during infection can be determined. We utilized cells that can be efficiently infected by VSV-G-pseudotyped retroviruses, and fractionated the cytosolic proteins on linear gradients following synchronized infection. The fates of capsid and integrase proteins, as well as viral genomic RNA and reverse transcription products were then monitored. We found that components of MLV and HIV-1 cores formed a large complex under non-restrictive conditions. In contrast, when MLV infection was restricted by human TRIM5α, the integrase protein and reverse transcription products were lost from infected cells, while capsid and viral RNA were both solubilized. Similarly, when HIV-1 infection was restricted by rhesus TRIM5α or owl monkey TRIMCyp, the integrase protein and reverse transcription products were lost. However, viral RNA was also lost, and high levels of preexisting soluble CA prevented the determination of whether CA was solubilized. Notably, proteasome inhibition blocked all of the aforementioned biochemical consequences of TRIM5α-mediated restriction but had no effect on its antiviral potency. Together, our results show how TRIM5α affects various retroviral core components and indicate that proteasomes are required for TRIM5α-induced core disruption but not for TRIM5α-induced restriction.
format	article
author	Sebla B Kutluay David Perez-Caballero Paul D Bieniasz
author_facet	Sebla B Kutluay David Perez-Caballero Paul D Bieniasz
author_sort	Sebla B Kutluay
title	Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
title_short	Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
title_full	Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
title_fullStr	Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
title_full_unstemmed	Fates of retroviral core components during unrestricted and TRIM5-restricted infection.
title_sort	fates of retroviral core components during unrestricted and trim5-restricted infection.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/273780abbffe4f918789c7230f890c8b
work_keys_str_mv	AT seblabkutluay fatesofretroviralcorecomponentsduringunrestrictedandtrim5restrictedinfection AT davidperezcaballero fatesofretroviralcorecomponentsduringunrestrictedandtrim5restrictedinfection AT pauldbieniasz fatesofretroviralcorecomponentsduringunrestrictedandtrim5restrictedinfection
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Fates of retroviral core components during unrestricted and TRIM5-restricted infection.

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