September 2009 Meeting Announcement, Delaware Valley Mass Spectrometry Discussion Group
PLEASE NOTE ROOM CHANGE: We will meet in Mendel 102.
- Topic: "Targeted Proteomic Approaches Provide Insights into Virion Assembly and Chromatin Remodeling during Viral Infection
- Speaker: Ileana Cristea, Princeton University
- Date: Monday, September 14, 2009. 6:30 PM
- Time: Social Hour: 6:30 PM.
Talk: 7:30 PM.
Please RSVP to John Masucci JMASUCCI@PRDUS.JNJ.COM by Thursday September 10th.
- Place: Department of Chemistry, Villanova University (Room 102, Mendel Hall)
Viruses have evolved finely tuned interactions with their hosts to manipulate and adapt complex cellular processes for their own use. The study of virus-host interactions has therefore emerged as a key driving force in the research of infectious disease during the post-genomic era. Despite these efforts, our understanding of the protein interactome remains, in large part, unknown. The development and incorporation of new approaches that can reveal the dynamics of virus-host protein interactions is a necessity. Modern proteomic techniques have the ability to provide access to such interactions, and the ever increasing sensitivity of mass spectrometry allows the identification and quantification of relatively low levels of proteins. This presentation will describe targeted proteomic approaches for studying virus-host macromolecular assemblies. Highlights will be shown from our studies on infections with human immunodeficiency virus (HIV) and human cytomegalovirus (HCMV).
We employed targeted genetic-proteomic approaches to study the virus-host interface either from the virus or host perspective. Using a library of tagged replication competent HCMV and HIV mutant viruses, we infected primary human fibroblasts (for HCMV) and CEM T cells (for HIV), and employed cryogenic cell lysis and rapid immunoaffinity purifications on magnetic beads to isolate virus-host assemblies. For studies on histone deacetylases (HDAC) during viral infections, we generated cell lines stably expressing green fluorescent protein tagged HDACs and probed their interactions and deacetylation activity. Isolated protein complexes were analyzed using a MALDI LTQ Orbitrap (Thermo Fisher Scientific) and the specificity of observed interactions was confirmed by immunofluorescence, reciprocal immunoprecipitation and metabolic labeling with stable isotopes (I-DIRT).
Two interesting findings will be highlighted: 1) studies on pUL32, pUL99, pUL83 and pTRS1 HCMV proteins demonstrated that parallel processes occur at distinct cellular sites during the assembly of HCMV virions, and 2) chromatin remodeling complexes, including histone deacetylases, are targeted by viruses, possibly in part to gain control over host gene expression and modulate the outcome of an infection.
Ileana Cristea has performed her graduate research at the Michael Barber Center for Mass Spectrometry, University of Manchester, U.K., under the supervision of Simon Gaskell, and at the Toxicology Research & Development Department at GlaxoSmithKline, U.K. She then pursued her postdoctoral work in the mass spectrometry laboratory of Brian Chait at The Rockefeller University. She is currently an Assistant Professor in the Department of Molecular Biology at Princeton University, where her laboratory focuses on understanding mechanisms that control the fate of cells under invasion by pathogens, with particular emphasis on the viral modulation of chromatin remodeling machineries. Dr. Cristea is part of the American Society for Mass Spectrometry Education Committee. Her teaching activities have included a section of the Cold Spring Harbor Proteomics Course focused on isolating protein complexes (2006-2009). She is the recipient of numerous awards, including the Bordoli Prize from the British Mass Spectrometry Society (2001), NIDA Avant-Garde Award for HIV/AIDS Research (2008), and Human Frontiers Science Program Young Investigator Grant Award (2009).
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