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Proteomics of HCMV


EMSL Project ID
12295

Abstract

The HCMV project is an NCRR Biomedical Technology Resource Center project. The main aim of this proposal is to develop and exploit new approaches for comprehensive proteome measurements that will serve as the basis for identifying human cytomegalovirus (HCMV) proteins and associated host proteins. HCMV, a member of the herpesviruses, is one of the most common opportunistic pathogens encountered in patients immuno-compromised from congenital or acquired diseases, such as AIDS, or through transplantation procedures. HCMV is a large complex enveloped virus and encodes over 200 predicted ORFs. While the sequence of the HCMV genome has been known for over a decade, the full set of viral and cellular proteins that compose the HCMV virion are unknown. Previous work from our group has utilized ‘gel-free’ 2-D capillary liquid chromatography (LC)-MS/MS and Fourier transform ion cyclotron resonance (FTICR) mass spectrometry to identify and determine the relative abundances of viral and cellular proteins in purified HCMV AD169 virions and dense bodies. We identified 71 HCMV encoded proteins that include 12 proteins encoded by known viral ORFs previously not associated with virions and 12 proteins from novel viral ORFs. We have also identified over 75 host cellular proteins in HCMV virions that include cellular structural proteins, enzymes, and chaperones. We would like to extend these studies to increase our understanding of the events occuring during HCMV infection, through a comparison of the viral proteome to the viral transcriptome measured under the same conditions; specifically to compare the time course of changes in viral mRNA expression over the course of infection with the appearance and persistence of the protein product. Additionally, in order to increase our understanding of the virus’ ability to escape the host immune response, we propose an analysis of the specific protein-protein complexes formed between the host membrane proteins and the viral proteins. Further studies will investigate the differences between laboratory strains and clinically isolated strains of HCMV.

Project Details

Project type
Exploratory Research
Start Date
2005-02-24
End Date
2007-09-02
Status
Closed

Team

Principal Investigator

Susan Varnum
Institution
Pacific Northwest National Laboratory

Team Members

Jon Jacobs
Institution
Environmental Molecular Sciences Laboratory

Related Publications

Dumortier J, Streblow DN, Moses AV, Jacobs JM, Kreklywich CN, Camp D, Smith RD, Orloff SL, Nelson JA: Human cytomegalovirus secretome contains factors that induce angiogenesis and wound healing; J Virol. 2008 Jul;82(13):6524-35.
Prichard MN, Sztul E, Daily SL, Perry AL, Frederick SL, Gill RB, Hartline CB, Streblow DN, Varnum SM, Smith RD, Kern ER.Human cytomegalovirus UL97 kinase activity is required for the hyperphosphorylation of retinoblastoma protein and inhibits the formation of nuclear aggresomes. J. Virol. (2008) 82: 5054-5067
Streblow DN, Varnum SM, Smith RD, and J Nelson. (2006)."A Proteomics Analysis of Human Cytomegalovirus Particles." In Cytomegaloviruses: Pathogenesis, Molecular Biology, and Infection Control. Edited by Matthias Reddehase. Caister Academic Press, Norwich, England.
Varnum SM, Streblow DN, Monroe ME, Smith P, Auberry KJ, Pasa-Tolic L, Wang D, Camp DG 2nd, Rodland K, Wiley S, Britt W, Shenk T, Smith RD, Nelson JA. Identification of proteins in human cytomegalovirus (HCMV) particles: the HCMV proteome. J Virol. 2004 Oct;78(20):10960-6.