Genetic and Functional Dissection of the Role of Individual 5-HT2 Receptors as Entry Receptors for JC Polyomavirus.

Abstract

JC polyomavirus (JCPyV) is a ubiquitous human pathogen that causes progressive multifocal leukoencephalopathy (PML). The entry receptors for JCPyV belong to the 5-hydroxytryptamine 2 receptor (5-HT2R) family, but how individual members of the family function to facilitate infection is not known. We used proximity ligation assay (PLA) to determine that JCPyV interacts with each of the 5-HT2 receptors (5-HT2Rs) in a narrow window of time during entry. We used CRISPR-Cas9 to randomly introduce stop codons in the gene for each receptor and discovered that the second intracellular loop of each was necessary for infection. This loop contains a motif possibly involved in receptor internalization by β-arrestin. Mutation of this motif and small interfering RNA (siRNA) knockdown of β-arrestin recapitulated the results of our CRISPR-Cas9 screen, showing that this motif is critical. Our results have implications for the role these receptors play in virus infection and for their normal functioning as receptors for serotonin.

• Authors: Benedetta Assetta, Jenna Morris-Love, Gretchen V Gee, Abigail L Atkinson, Bethany A O'Hara, Melissa S Maginnis, Sheila A Haley, Walter J Atwood
• Journal: Cell reports (Cell Rep) Vol. 27 Issue 7 Pg. 1960-1966.e6 (05 14 2019) ISSN: 2211-1247 [Electronic] United States
• PMID: 31091436 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Receptors, Serotonin, 5-HT2
• Receptors, Virus
• beta-Arrestins

Topics

• HEK293 Cells
• Host-Pathogen Interactions (genetics)
• Humans
• JC Virus (genetics, pathogenicity)
• Receptors, Serotonin, 5-HT2 (genetics, metabolism)
• Receptors, Virus (genetics, metabolism)
• Virus Internalization
• beta-Arrestins (genetics, metabolism)