Talk Title: TMT-based proteomic analysis identifies novel viral evasion pathways
Paul Lehner trained in medicine and infectious diseases in London and currently holds a Wellcome Trust Principal Research Fellowship at the CIMR, University of Cambridge. He uses novel functional genetic and proteomic technologies to study how viruses interact with, and evade recognition by, the human immune system. His aims are to identify cellular receptors manipulated by viruses and understand how and why these receptors are targeted. His group used unbiased functional SILAC/TMT-based proteomic approaches, called plasma membrane profiling, to identify novel cell surface receptors manipulated by viruses. More recently he has applied systematic unbiased TMT-based proteomics to determine how viral infections (eg HIV, HCMV) affect the whole cell proteome Complementary to this approach is the use of genome-wide fluorescent-based genetic selection screens, to map genetic pathways which regulate receptor expression. Together these technologies provide a protein and gene discovery platform to identify novel genes and pathways required for receptor regulation.
TMT-based proteomic analysis identifies novel viral evasion pathways
Evolutionary pressure has produced an “arms race” between cellular restriction factors that limit viral replication and viral proteins which overcome host restriction. Thus viruses need to manipulate host cell signaling pathways to enable their replication and evade immune recognition. In turn, infected cells must sense and respond appropriately to intracellular viral infections. Many virus-induced changes in the host proteome are post-translational and therefore lend themselves to proteomic analysis. We have adapted functional proteomic approaches to identify receptors manipulated by viruses and identify key components of intracellular signaling pathways. We applied ‘Plasma Membrane Profiling’ to gain a systematic unbiased overview of cell surface receptors manipulated by viruses, as well as TMT-based proteomic analysis to delineate virus-induced changes in the total cell proteome. TMT-based quantitation allows us to create a dynamic temporal map of receptors manipulated by both integrating (HIV) and non-integrating (herpes) viruses. This approach has established novel paradigms of viral interference and manipulation of the host immune system which will be discussed.