Quantitative proteomic analysis of latent human cytomegalovirus infection (360G-Wellcome-093966_Z_10_Z)

£243,917

Following primary infection, HCMV typically establishes a latent infection in monocytes under the control of a healthy immune system. Little is known about the intracellular pathways manipulated by HCMV to maintain latency, or the phenotype of latently infected cells. I will: (i) identify cell surface targets of individual HCMV latency genes (ii) enrich ex vivo latently infected monocytes to characterise the most important biochemical pathways altered by latent HCMV infection. I have developed a new technique ?plasma membrane profiling? (PMP) to enrich plasma membrane (PM) proteins for proteomic analysis. I will employ the functional proteomics approach SILAC (Stable Isotope Labelling by Amino acids in Cell culture), to perform semi-quantitative analysis on plasma membrane proteins enriched by PMP. Using this approach I will determine how latent HCMV infection alters the cell surface proteome by analysing transduced cell lines expressing individual HCMV latency genes, and an in vitro model of latency. I shall use the markers I identify to enrich latently-infected ex vivo CD34+ monocytes. In addition, I will characterise biochemical pathways altered by latent HCMV infection in these cells using ?KAYAK? (Kinase ActivitY Assay for Kinome profiling), a novel proteomic technique. Candidate proteomic ?hits? will be biochemically characterised.

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Grant Details

Amount Awarded 243917
Applicant Surname Weekes
Approval Committee Clinical Interview Committee
Award Date 2010-11-25T00:00:00+00:00
Financial Year 2010/11
Grant Programme: Title Postdoctoral Training Fellowship for Clinicians
Internal ID 093966/Z/10/Z
Lead Applicant Dr Michael Weekes
Partnership Value 243917
Planned Dates: End Date 2014-07-03T00:00:00+00:00
Planned Dates: Start Date 2011-07-04T00:00:00+00:00
Recipient Org: Country United Kingdom
Region East of England
Sponsor(s) Prof Paul Lehner