Proteomic identification of nuclear processes manipulated by cytomegalovirus early during infection. Proteomics 2015 Jun;15(12):1995-2005
Date
03/12/2015Pubmed ID
25758553Pubmed Central ID
PMC4556115DOI
10.1002/pmic.201400599Scopus ID
2-s2.0-84931005724 (requires institutional sign-in at Scopus site) 10 CitationsAbstract
Human cytomegalovirus (HCMV) is a herpesvirus that is ubiquitously distributed worldwide and causes life-threating disease upon immunosuppression. HCMV expresses numerous proteins that function to establish an intracellular environment that supports viral replication. Like most DNA viruses, HCMV manipulates processes within the nucleus. We have quantified changes in the host cell nuclear proteome at 24 h post infection following infection with a clinical viral isolate. We have combined SILAC with multiple stages of fractionation to define changes. Tryptic peptides were analyzed by RP-HPLC combined with LC-MS/MS on an LTQ Orbitrap Velos mass spectrometer. Data from three biological replicates were processed with MaxQuant. A total of 1281 cellular proteins were quantified and 77 were found to be significantly differentially expressed. In addition, we observed 36 viral proteins associated with the nucleus. Diverse biological processes were significantly altered, including increased aspects of cell cycling, mRNA metabolism, and nucleocytoplasmic transport and decreased immune responses. We validated changes for several proteins including a subset of classical nuclear transport proteins. In addition, we demonstrated that disruption of these import factors is inhibitory to HCMV replication. Overall, we have identified HCMV-induced changes in the nuclear proteome and uncovered several processes that are important for infection. All MS data have been deposited in the ProteomeXchange with identifier PXD001909 (http://proteomecentral.proteomexchange.org/dataset/PXD001909).
Author List
Carter DM, Westdorp K, Noon KR, Terhune SSAuthor
Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Blotting, WesternCell Nucleus
Cells, Cultured
Chromatography, Liquid
Cytomegalovirus
Cytomegalovirus Infections
Fibroblasts
Humans
Immunoprecipitation
Nuclear Proteins
Proteomics
Tandem Mass Spectrometry
Viral Proteins
Virus Replication