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Proteomic identification of nuclear processes manipulated by cytomegalovirus early during infection. Proteomics 2015 Jun;15(12):1995-2005

Date

03/12/2015

Pubmed ID

25758553

Pubmed Central ID

PMC4556115

DOI

10.1002/pmic.201400599

Scopus ID

2-s2.0-84931005724   7 Citations

Abstract

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 SS

Author

Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Blotting, Western
Cell Nucleus
Cells, Cultured
Chromatography, Liquid
Cytomegalovirus
Cytomegalovirus Infections
Fibroblasts
Humans
Immunoprecipitation
Nuclear Proteins
Proteomics
Tandem Mass Spectrometry
Viral Proteins
Virus Replication