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http://hungrydogweb.com/?p=47975 Introduction: HAUSP, Herpesvirus-Associated Ubiquitin-Specific Protease, a deubiquitinating enzyme, has varied roles in a number of biological processes ranging from genome stability, epigenetic regulation, cell cycle and apoptosis, to viral infection, immunity and stem cell maintenance and hence, emerges to be a very important candidate with implications in cancer as well as other pathologies. The dual, context-specific role of HAUSP, both as an oncogene and a tumor suppressor, makes its role versatile. Materials and Methods : HAUSP was cloned in pGZ vector and transfected in HEK293 cells. Whole cell lysates from vector and HAUSP transfected cells were separated by 2D-gel electrophoresis. Gels were coomassie stained and spots were identified using PDQuest software. Spots were excised and tryptic digests of proteins isolated using In-gel trypsinization kit. MALDI-TOF/TOF MS/MS and GPS analysis were performed for identification of proteins. Immunoprecipitates from HAUSP overexpressed cells were analyzed similarly for identification of putative interacting partners of HAUSP. Results and Conclusion: In this study, we have shown the role of exogenously expressed HAUSP on the overall proteome. Of the 38 proteins identified, 16 were upregulated, 8 down-regulated, 7 unique spots and 3 spots were absent when compared to the control gel. The proteins identified indicate a probable involvement of HAUSP in tumor suppression, transport, metabolism, cytoskeletal organization, gene expression, chaperones, apoptosis etc. Enriched pool of interacting proteins by immunoprecipitation has elucidated a number of probable and yet unknown partners. Of the 36 interacting proteins, some of the proteins like TRRAP, FBF-1, etc. with changed profiles also interact with HAUSP. Gene ontology based classification of the proteins reveals several enzymes, proteins involved in transportation, apoptosis, gene expression, etc. Five Zinc Finger Proteins were identified with the same C2H2 domain. Also a few uncharacterized proteins have been identified. The overall aim of this study is to explore the global picture of the changes in the protein profile and putative interacting partners of HAUSP by mass spectrometric analysis, hence attributing newer functions to the protein. Funniest weed joke? What You Need to Roll a Blunt.
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