title: Proteomics analysis of herpes simplex virus type 1-infected cells reveals dynamic changes of viral protein expression, ubiquitylation, and phosphorylation.
abstract: Herpesviruses are among the most complex and widespread human viruses and cause a number of diseases ranging from cold sores to genital infections and encephalitis. While the composition of viral particles has been studied, less is known about the expression of the whole viral proteome in infected cells. Here, we analyzed the proteome of the prototypical Herpes Simplex Virus type 1 (HSV1) in infected cells by mass spectrometry. Using a high sensitivity LTQ-Orbitrap, we achieved a very high level of protein coverage and identified a total of 67 structural and nonstructural viral proteins. We also identified 90 novel phosphorylation sites and 10 novel ubiquitylation sites on different viral proteins. Ubiquitylation was observed on nine HSV1 proteins. We identified phosphorylation sites on about half of the detected viral proteins; many of the highly phosphorylated ones are known to regulate gene expression. Treatment with inhibitors of DNA replication induced changes of both viral protein abundance and modifications, highlighting the interdependence of viral proteins during the life cycle. Given the importance of expression dynamics, ubiquitylation, and phosphorylation for protein function, these findings will serve as important tools for future studies on herpesvirus biology.
authors: Département de Chimie, ‡Proteomics and Mass Spectrometry Research Unit, Institute for Research in Immunology and Cancer, and §Département de Pathologie et Biologie Cellulaire, Université de Montréal, C.P. 6128-Succursale Centre-Ville, Montréal, Québec H3C 3J7, Canada.
authors: Christina
authors: C
authors: Bell
authors: Michel
authors: M
authors: Desjardins
authors: Pierre
authors: P
authors: Thibault
authors: Kerstin
authors: K
authors: Radtke
mesh_terms: D000212:Acyclovir / Q000494:pharmacology; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line / Q000821:virology; D003513:Cycloheximide / Q000494:pharmacology; D004261:DNA Replication / Q000187:drug effects; D015967:Gene Expression Regulation, Viral; D018259:Herpesvirus 1, Human / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000821:virology; D051379:Mice; D008810:Mice, Inbred C57BL; D010746:Phosphonoacetic Acid / Q000494:pharmacology; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D040901:Proteomics / Q000379:methods; D054875:Ubiquitination; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D003513:Cycloheximide; D010746:Phosphonoacetic Acid; D000212:Acyclovir