Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion

ABSTRACT Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the Old World and a highly fatal hantavirus cardiopulmonary syndrome (HCPS) in the New World. No vaccines or antiviral therapies are currently available to prevent or treat hantavirus disease, and gaps in our understanding o...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Lara M. Kleinfelter, Rohit K. Jangra, Lucas T. Jae, Andrew S. Herbert, Eva Mittler, Katie M. Stiles, Ariel S. Wirchnianski, Margaret Kielian, Thijn R. Brummelkamp, John M. Dye, Kartik Chandran
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2015
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/983bfed9990349deb89d2c2be0fb9944
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:983bfed9990349deb89d2c2be0fb9944
record_format	dspace
spelling	oai:doaj.org-article:983bfed9990349deb89d2c2be0fb99442021-11-15T15:41:27ZHaploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion10.1128/mBio.00801-152150-7511https://doaj.org/article/983bfed9990349deb89d2c2be0fb99442015-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00801-15https://doaj.org/toc/2150-7511ABSTRACT Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the Old World and a highly fatal hantavirus cardiopulmonary syndrome (HCPS) in the New World. No vaccines or antiviral therapies are currently available to prevent or treat hantavirus disease, and gaps in our understanding of how hantaviruses enter cells challenge the search for therapeutics. We performed a haploid genetic screen in human cells to identify host factors required for entry by Andes virus, a highly virulent New World hantavirus. We found that multiple genes involved in cholesterol sensing, regulation, and biosynthesis, including key components of the sterol response element-binding protein (SREBP) pathway, are critical for Andes virus entry. Genetic or pharmacological disruption of the membrane-bound transcription factor peptidase/site-1 protease (MBTPS1/S1P), an SREBP control element, dramatically reduced infection by virulent hantaviruses of both the Old World and New World clades but not by rhabdoviruses or alphaviruses, indicating that this pathway is broadly, but selectively, required by hantaviruses. These results could be fully explained as arising from the modest depletion of cellular membrane cholesterol that accompanied S1P disruption. Mechanistic studies of cells and with protein-free liposomes suggested that high levels of cholesterol are specifically needed for hantavirus membrane fusion. Taken together, our results indicate that the profound dependence on target membrane cholesterol is a fundamental, and unusual, biophysical property of hantavirus glycoprotein-membrane interactions during entry. IMPORTANCE Although hantaviruses cause important human diseases worldwide, no specific antiviral treatments are available. One of the major obstacles to the development of new therapies is a lack of understanding of how hantaviruses hijack our own host factors to enter cells. Here, we identified multiple cellular genes that control the levels of cholesterol in cellular membranes to be important for hantavirus entry. Our findings suggest that high concentrations of cholesterol in cellular membranes are required at a specific step in the entry process—fusion between viral and cellular membranes—that allows escape of the hantavirus genome into the host cell cytoplasm to initiate infection. Our findings uncover a fundamental feature of the hantavirus infection mechanism and point to cholesterol-lowering drugs as a potential new treatment of hantaviral infections.Lara M. KleinfelterRohit K. JangraLucas T. JaeAndrew S. HerbertEva MittlerKatie M. StilesAriel S. WirchnianskiMargaret KielianThijn R. BrummelkampJohn M. DyeKartik ChandranAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 6, Iss 4 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Lara M. Kleinfelter Rohit K. Jangra Lucas T. Jae Andrew S. Herbert Eva Mittler Katie M. Stiles Ariel S. Wirchnianski Margaret Kielian Thijn R. Brummelkamp John M. Dye Kartik Chandran Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
description	ABSTRACT Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the Old World and a highly fatal hantavirus cardiopulmonary syndrome (HCPS) in the New World. No vaccines or antiviral therapies are currently available to prevent or treat hantavirus disease, and gaps in our understanding of how hantaviruses enter cells challenge the search for therapeutics. We performed a haploid genetic screen in human cells to identify host factors required for entry by Andes virus, a highly virulent New World hantavirus. We found that multiple genes involved in cholesterol sensing, regulation, and biosynthesis, including key components of the sterol response element-binding protein (SREBP) pathway, are critical for Andes virus entry. Genetic or pharmacological disruption of the membrane-bound transcription factor peptidase/site-1 protease (MBTPS1/S1P), an SREBP control element, dramatically reduced infection by virulent hantaviruses of both the Old World and New World clades but not by rhabdoviruses or alphaviruses, indicating that this pathway is broadly, but selectively, required by hantaviruses. These results could be fully explained as arising from the modest depletion of cellular membrane cholesterol that accompanied S1P disruption. Mechanistic studies of cells and with protein-free liposomes suggested that high levels of cholesterol are specifically needed for hantavirus membrane fusion. Taken together, our results indicate that the profound dependence on target membrane cholesterol is a fundamental, and unusual, biophysical property of hantavirus glycoprotein-membrane interactions during entry. IMPORTANCE Although hantaviruses cause important human diseases worldwide, no specific antiviral treatments are available. One of the major obstacles to the development of new therapies is a lack of understanding of how hantaviruses hijack our own host factors to enter cells. Here, we identified multiple cellular genes that control the levels of cholesterol in cellular membranes to be important for hantavirus entry. Our findings suggest that high concentrations of cholesterol in cellular membranes are required at a specific step in the entry process—fusion between viral and cellular membranes—that allows escape of the hantavirus genome into the host cell cytoplasm to initiate infection. Our findings uncover a fundamental feature of the hantavirus infection mechanism and point to cholesterol-lowering drugs as a potential new treatment of hantaviral infections.
format	article
author	Lara M. Kleinfelter Rohit K. Jangra Lucas T. Jae Andrew S. Herbert Eva Mittler Katie M. Stiles Ariel S. Wirchnianski Margaret Kielian Thijn R. Brummelkamp John M. Dye Kartik Chandran
author_facet	Lara M. Kleinfelter Rohit K. Jangra Lucas T. Jae Andrew S. Herbert Eva Mittler Katie M. Stiles Ariel S. Wirchnianski Margaret Kielian Thijn R. Brummelkamp John M. Dye Kartik Chandran
author_sort	Lara M. Kleinfelter
title	Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
title_short	Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
title_full	Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
title_fullStr	Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
title_full_unstemmed	Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion
title_sort	haploid genetic screen reveals a profound and direct dependence on cholesterol for hantavirus membrane fusion
publisher	American Society for Microbiology
publishDate	2015
url	https://doaj.org/article/983bfed9990349deb89d2c2be0fb9944
work_keys_str_mv	AT laramkleinfelter haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT rohitkjangra haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT lucastjae haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT andrewsherbert haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT evamittler haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT katiemstiles haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT arielswirchnianski haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT margaretkielian haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT thijnrbrummelkamp haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT johnmdye haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion AT kartikchandran haploidgeneticscreenrevealsaprofoundanddirectdependenceoncholesterolforhantavirusmembranefusion
_version_	1718427718109888512

Haploid Genetic Screen Reveals a Profound and Direct Dependence on Cholesterol for Hantavirus Membrane Fusion

Ejemplares similares