91丝瓜视频

Dr. Anne-Claude Gingras
Assistant Professor
Department of Molecular Genetics
University of Toronto

鈥淐harting kinase and phosphatase interactomes: From interaction proteomics to functional insights鈥�

Anne-Claude Gingras received her Ph.D. from 91丝瓜视频 University in 2001 for her studies on translational control, performed under the guidance of Nahum Sonenberg.听 She then joined the laboratory of Ruedi Aebersold at the Institute for Systems Biology in Seattle, where she developed proteomics approaches to study protein-protein interactions.听 She established her own laboratory in 2005 at the Samuel Lunenfeld Research Institute at Mount Sinai Hospital in Toronto, where she currently holds a Canada Research Chair in Functional Proteomics and the Lea Reichmann Chair in Cancer Proteomics.听 She is an Assistant Professor in the Department of Molecular Genetics of the University of Toronto. Her group focuses on the identification and functional characterization of protein interaction modules surrounding serine/threonine phosphatases. Active areas of research include understanding how the PP4 phosphatase modulates gene expression via regulation of transcription elongation, mRNA capping, and splicing. The Gingras lab is also investigating the function of a novel protein complex that they have identified surrounding the protein CCM3, whose gene is mutated in familial cases of angioma.听 Anne-Claude Gingras is also involved in the development of robust approaches for the analysis of protein interaction networks in yeast and human and for the study of enzyme-substrate relationships within the interaction networks.

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Protein phosphorylation mediates cellular responses to growth factors, environmental signals, and internal processes by the regulation of protein interactions, enzyme activity or protein localization. However, the protein interactions of kinases, phosphatases, their regulatory subunits and substrates remain sparsely mapped. Two interaction mapping projects will be presented: 1) To chart the budding yeast kinase and phosphatase interaction (KPI) network, we systematically characterized protein kinase and phosphatase complexes by sensitive affinity purification coupled to mass spectrometric identification (AP-MS).听 We identified a KPI network of 1,844 interactions that contained many dense local regions.听 Notably, the cell cycle phosphatase Cdc14 associated with multiple kinases that revealed roles for Cdc14 in mitogen-activated protein kinase signaling, the DNA damage response and metabolism, while interactions of the target of rapamycin complex 1 (TORC1) uncovered new effector kinases in nitrogen and carbon metabolism. An extensive backbone of kinase-kinase interactions cross-connects the proteome and may serve to coordinate diverse cellular responses. 2) To identify potential new substrates, regulators and targeting subunits for the human PP2A phosphatase, we performed an iterative AP-MS approach.听 This allowed us to uncover an interaction network containing 365 proteins.听 Within this network, we further focused on the characterization of the largest stable PP2A containing complex identified to date.听 This complex, called STRIPAK, is evolutionarily conserved and implicated in cytoskeletal events.听 A core component of STRIPAK, CCM3, is mutated in familial cases of cerebral cavernous malformations, which are vascular anomalies of the brain capillaries, and our recent results indicate that STRIPAK is involved in the disease.听 These two projects will be presented within the context of the development of bioinformatics and statistical analysis tools for protein-protein interactions.

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General review of interaction proteomics:

Gingras et al., Nat Review Mol Cell Biol, 2007, 8:645

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Protocols for affinity purification coupled to mass spectrometry

Chen and Gingras, Methods, 2007, 42-298

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The scientific presentation will focus on unpublished data and:

1) Goudreault et al., Mol Cell Proteomics, 2009, 8:157

2) Breitkreutz et al., Science, 2010 (in press)

3) Choi et al., Mol Sys Biol, 2010 (in press)