Role of PDGF receptor-α during human cytomegalovirus entry into fibroblasts.

TitleRole of PDGF receptor-α during human cytomegalovirus entry into fibroblasts.
Publication TypeJournal Article
Year of Publication2018
AuthorsWu, K, Oberstein, A, Wang, W, Shenk, T
JournalProc Natl Acad Sci U S A
Volume115
Issue42
PaginationE9889-E9898
Date Published2018 10 16
ISSN1091-6490
Abstract

Human CMV (HCMV) exhibits a broad cell tropism that depends on two virion glycoprotein complexes: a trimeric complex (gH/gL/gO) that facilitates viral infection primarily in fibroblasts and a pentameric complex (gH/gL/pUL128-pUL130-pUL131A) that mediates infection in epithelial and endothelial cells. We performed genome-wide CRISPR screens in which the PDGF receptor-α (PDGFRα) was identified as the most significant cellular gene product essential for infection by HCMV virions containing only trimeric complex (trimer-only virus). Trimer-only virus did not enter PDGFRα knockout fibroblasts. By using knockout fibroblasts, the extracellular domain of PDGFRα required for virus entry was mapped, and the intracellular tyrosine kinase domain was shown to be nonessential. In addition, direct cell-to-cell spread of virus from knockout cells transfected with trimer-only viral DNA was blocked, despite the production of infectious virus in the transfected cells. In contrast to trimer-only virus, wild-type HCMV virions containing both trimeric and pentameric complexes entered PDGFRα knockout cells, reinforcing the view that fibroblasts contain a second, independent receptor for the pentameric complex. Importantly, however, wild-type virus entered the knockout fibroblasts at reduced efficiency compared with parental fibroblasts, arguing that the cellular receptor for the virion pentameric complex is limiting or that virions are produced containing different relative amounts of the two glycoprotein complexes. Finally, ectopic expression of PDGFRα in ARPE-19 epithelial cells and THP-1 monocytic cells, which have little to no endogenous PDGFRα expression, markedly enhanced their susceptibility to trimer-only virions. In sum, our data clarify several key determinants of HCMV tropism.

DOI10.1073/pnas.1806305115
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID30275317
PubMed Central IDPMC6196492
Grant ListF32 AI106175 / AI / NIAID NIH HHS / United States
R01 AI112951 / AI / NIAID NIH HHS / United States