Cited by 40SARS-CoV-2 and COVID-19 ResearchAdvanced Proteomics Techniques and ApplicationsAnimal Virus Infections Studies

Identification of host factors involved in coronavirus replication by quantitative proteomics analysis

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Vogels, Mijke W., van Balkom, Bas W. M., Kaloyanova, Dora V., Batenburg, Joseph J., Heck, Albert J., Helms, J. Bernd, Rottier, Peter J. M., de Haan, Cornelis A. M.·2011·PROTEOMICS

Abstract

In this study, we applied a quantitative proteomic approach, based on SILAC, to investigate the interactions of coronaviruses with the secretory pathway of the host cell, with the aim to identify host factors involved in coronavirus replication. Comparison of the protein profiles of Golgi‐enriched fractions of cells that were either mock infected or infected with mouse hepatitis virus revealed the significant depletion or enrichment of 116 proteins. Although ribosomal/nucleic acid binding proteins were enriched in the Golgi‐fractions of mouse hepatitis virus‐infected cells, proteins annotated to localize to several organelles of the secretory pathway were overrepresented among the proteins that were depleted from these fractions upon infection. We hypothesized that proteins, of which the abundance or distribution is affected by infection, are likely to be involved in the virus life cycle. Indeed, depletion of a small subset of the affected proteins by using small interfering RNAs identified several host factors involved in coronavirus infection. Transfection of small interfering RNAs targeting either C11orf59 or Golgi apparatus glycoprotein 1 resulted in increased virus replication, whereas depletion of vesicle‐trafficking protein vesicle‐trafficking protein sec22b enhanced the release of infectious progeny virus. Overexpression of these proteins, on the other hand, had a negative effect on virus replication. Overall, our study shows that the SILAC approach is a suitable tool to study host–pathogen interactions and to identify host proteins involved in virus replication.

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  • Differentially regulated Golgi-resident and vesicle trafficking factors, including SEC22B and LAMTOR1, were shown to affect MHV virion assembly in RNAi and overexpression assays.

    “selected differentially regulated Golgi-resident and vesicle trafficking factors, such as SEC22B and the Ragulator complex protein LAMTOR1, were shown to alter MHV virion assembly using RNA interference and overexpression assays.”
  • In mouse hepatitis virus-infected hepatocytes, SILAC measured virus-induced expression changes in Golgi-enriched fractions.

    “metabolic labeling using stable isotope labeling in cell culture (SILAC; see Figure 2) was employed to quantify virus-induced expression changes in Golgi-enriched fractions from mouse hepatitis virus (MHV)-infected hepatocytes (102).”

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Authors
Vogels, M. W., van Balkom, B. W. M., Kaloyanova, D. V., Batenburg, J. J., Heck, A. J., Helms, J. B., Rottier, P. J. M., & de Haan, C. A. M.
Affiliations
Utrecht University (NL)
Venue
PROTEOMICS
Publisher
Wiley
Published
2010-12-06
Volume
11
Issue
1
Pages
64-80
ISSN
  • 1615-9853 (ISSN-L)
  • 1615-9861
OpenAlex
W2040208749
Cited by
40
References
69
Citations by year
2020: 2 · 2019: 1 · 2018: 1 · 2017: 1 · 2016: 5 · 2015: 4 · 2014: 6 · 2013: 3 · 2012: 10
FWCI
4.15
Type
journal article
Language
en
Open access
Locations
  • PROTEOMICS (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Utrecht University Repository (Utrecht University) (submittedVersion)
Funders
Faculty of Veterinary Medicine of Utrecht University, The Netherlands
Topics
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
Keywords
coronavirus replication, host factors, secretory pathway, Golgi apparatus, mouse hepatitis virus, host-pathogen interaction, Sec22b, vesicle trafficking, protein depletion, siRNA knockdown
MeSH terms
Electrophoresis, Polyacrylamide Gel, HeLa Cells, Murine hepatitis virus, Humans, Sialoglycoproteins, Mass Spectrometry, Virus Replication, Receptors, Fibroblast Growth Factor, Coronavirus, RNA Interference, RNA, Small Interfering, Proteomics, Host-Pathogen Interactions

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