Haploid genetic screen reveals a profound and direct dependence on cholesterol for hantavirus membrane fusion

Lara M. Kleinfelter, Rohit Jangra, Lucas T. Jae, Andrew S. Herbert, Eva-Maria Mittler, Katie M. Stiles, Ariel S. Wirchnianski, Margaret Kielian, Thijn R. Brummelkamp, John M. Dye, Kartik Chandran

Research output: Contribution to journalArticle

39 Citations (Scopus)

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.

Original languageEnglish (US)
Article numbere00801-15
JournalmBio
Volume6
Issue number4
DOIs
StatePublished - Jun 30 2015

Fingerprint

Hantavirus
Membrane Fusion
Haploidy
Cholesterol
Membranes
Hantavirus Infections
Response Elements
Sterols
Antiviral Agents
Carrier Proteins
Rhabdoviridae
Alphavirus
Hemorrhagic Fever with Renal Syndrome
Therapeutics
Virus Internalization
Critical Pathways
Membrane Glycoproteins
Infection
Hypercholesterolemia
Liposomes

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Haploid genetic screen reveals a profound and direct dependence on cholesterol for hantavirus membrane fusion. / Kleinfelter, Lara M.; Jangra, Rohit; Jae, Lucas T.; Herbert, Andrew S.; Mittler, Eva-Maria; Stiles, Katie M.; Wirchnianski, Ariel S.; Kielian, Margaret; Brummelkamp, Thijn R.; Dye, John M.; Chandran, Kartik.

In: mBio, Vol. 6, No. 4, e00801-15, 30.06.2015.

Research output: Contribution to journalArticle

Kleinfelter, Lara M. ; Jangra, Rohit ; Jae, Lucas T. ; Herbert, Andrew S. ; Mittler, Eva-Maria ; Stiles, Katie M. ; Wirchnianski, Ariel S. ; Kielian, Margaret ; Brummelkamp, Thijn R. ; Dye, John M. ; Chandran, Kartik. / Haploid genetic screen reveals a profound and direct dependence on cholesterol for hantavirus membrane fusion. In: mBio. 2015 ; Vol. 6, No. 4.
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