PXD023999

PXD023999 is an original dataset announced via ProteomeXchange.

Title	Viperin inhibits cholesterol biosynthesis and interacts with enzymes in the cholesterol biosynthetic pathway
Description	Many enveloped viruses bud from cholesterol-rich lipid rafts on the cell membrane. Depleting cellular cholesterol impedes this process and results in viral particles with reduced viability. Viperin (virus inhibitory protein endoplasmic reticulum-associated, interferon-induced) is an ER membrane-associated enzyme that when expressed in response to viral infections exerts broad-ranging antiviral effects, including inhibiting the budding of some enveloped viruses. Here we have investigated the effect of viperin expression on cholesterol biosynthesis. We found that viperin expression reduces cholesterol levels by 20 – 30 % in HEK293T cells. A proteomic screen of the viperin interactome identified several cholesterol biosynthetic enzymes among the top hits. The two most highly enriched proteins were lanosterol synthase and squalene monooxygenase, enzymes that catalyze key steps establishing the sterol carbon skeleton. Co-immunoprecipitation experiments established that viperin, lanosterol synthase and squalene monooxygenase form a complex at the ER membrane. Co-expression of viperin was found to significantly inhibit the specific activity of lanosterol synthase in HEK293T cell lysates. Co-expression of viperin had no effect on the specific activity of squalene monooxygenase, but reduced its expression levels in the cells by approximately 30 %. Despite these inhibitory effects, co-expression of either LS or SM failed to reverse the viperin-induced depletion of cellular cholesterol levels in HEK293T cells. Our results establish a clear link between the down-regulation of cholesterol biosynthesis and viperin, although at this point the effect cannot be unambiguously attributed interactions between viperin and a specific biosynthetic enzyme.
HostingRepository	PRIDE
AnnounceDate	2022-02-16
AnnouncementXML	Submission_2022-02-16_14:15:12.748.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Neil Marsh
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	ubiquitinylated lysine; phosphorylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-02-04 22:35:38	ID requested
⏵ 1	2022-02-16 14:15:13	announced

Grunkemeyer TJ, Ghosh S, Patel AM, Sajja K, Windak J, Basrur V, Kim Y, Nesvizhskii AI, Kennedy RT, Marsh ENG, The antiviral enzyme viperin inhibits cholesterol biosynthesis. J Biol Chem, 297(1):100824(2021) [pubmed]

submitter keyword: viperin

cholesterol regulation

interactome analysis

radical SAM enzyme

squalene monooxygenase

lanosterol synthase

Neil G Marsh
contact affiliation	Department of Chemistry, University of Michigan, Ann Arbor, MI 48109 Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109
contact email	nmarsh@umich.edu
lab head
Neil Marsh
contact affiliation	University of Michigan
contact email	nmarsh@umich.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/09/PXD023999

PRIDE project URI

• PRIDE
  1. PXD023999
     1. Label: PRIDE project
     2. Name: Viperin inhibits cholesterol biosynthesis and interacts with enzymes in the cholesterol biosynthetic pathway