Quantitative membrane proteomics reveals a role for tetraspanin enriched microdomains during entry of human cytomegalovirus

Kasinath Viswanathan, Marieke Verweij, Nessy John, Daniel Malouli, Klaus Frueh

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Human cytomegalovirus (HCMV) depends on and modulates multiple host cell membrane proteins during each stage of the viral life cycle. To gain a global view of the impact of HCMV-infection on membrane proteins, we analyzed HCMV-induced changes in the abundance of membrane proteins in fibroblasts using stable isotope labeling with amino acids (SILAC), membrane fractionation and protein identification by two-dimensional liquid chromatography and tandem mass spectrometry. This systematic approach revealed that CD81, CD44, CD98, caveolin-1 and catenin delta-1 were down-regulated during infection whereas GRP-78 was up-regulated. Since CD81 downregulation was also observed during infection with UV-inactivated virus we hypothesized that this tetraspanin is part of the viral entry process. Interestingly, additional members of the tetraspanin family, CD9 and CD151, were also downregulated during HCMV-entry. Since tetraspanin-enriched microdomains (TEM) cluster host cell membrane proteins including known CMV receptors such as integrins, we studied whether TEMs are required for viral entry. When TEMs were disrupted with the cholesterol chelator methyl-β-cylcodextrin, viral entry was inhibited and this inhibition correlated with reduced surface levels of CD81, CD9 and CD151, whereas integrin levels remained unchanged. Furthermore, simultaneous siRNA-mediated knockdown of multiple tetraspanins inhibited viral entry whereas individual knockdown had little effect suggesting essential, but redundant roles for individual tetraspanins during entry. Taken together, our data suggest that TEM act as platforms for receptors utilized by HCMV for entry into cells.

Original languageEnglish (US)
Article numbere0187899
JournalPLoS One
Volume12
Issue number11
DOIs
StatePublished - Nov 1 2017

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Human herpesvirus 5
Cytomegalovirus
Proteomics
proteomics
Membrane Proteins
membrane proteins
Membranes
Tetraspanins
integrins
Integrins
cell membranes
Down-Regulation
Isotope Labeling
Transmission electron microscopy
Caveolin 1
isotope labeling
receptors
Liquid chromatography
Cytomegalovirus Infections
Fibroblasts

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Quantitative membrane proteomics reveals a role for tetraspanin enriched microdomains during entry of human cytomegalovirus. / Viswanathan, Kasinath; Verweij, Marieke; John, Nessy; Malouli, Daniel; Frueh, Klaus.

In: PLoS One, Vol. 12, No. 11, e0187899, 01.11.2017.

Research output: Contribution to journalArticle

Viswanathan, Kasinath ; Verweij, Marieke ; John, Nessy ; Malouli, Daniel ; Frueh, Klaus. / Quantitative membrane proteomics reveals a role for tetraspanin enriched microdomains during entry of human cytomegalovirus. In: PLoS One. 2017 ; Vol. 12, No. 11.
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