University of California, Berkeley
Grant: $100,375 - National Institutes of Health - Sep. 15, 2009
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Award Description: Viral replication and immune evasion often require manipulation of cellular gene expression either in a targeted or global manner. Along these lines, lytic infection with the human gamma-herpesvirus Kaposi's sarcoma-associated herpesvirus (KSHV) leads to widespread inhibition of host gene expression. This phenotype is orchestrated by the KSHV SOX protein, which promotes accelerated degradation of mRNAs. SOX itself exhibits only weak ribonuclease activity, strongly suggesting that it must interface with one or more cellular pathways involved in RNA degradation. It is estimated that half of all gene expression is regulated at the level of RNA stability, yet most of these pathways remain poorly understood in human cells. The overall goals of this proposal are to characterize the mechanisms by which SOX promotes global mRNA destruction and establish what roles cellular factors play in this process. This will be accomplished through the following lines of experimentation: 1) systematic examination of the roles of established cellular mRNA turnover enzymes towards SOX activity, 2) development of an in vitro assay to monitor biochemically the mechanisms of RNA decay by SOX, and 3) identification of cellular proteins that interact with SOX. Funds provided by the Recovery Act will be used to hire a technician to assist in optimization of methodologies, generate of key reagents, and provide overall experimental support.
Project Description: The overall purpose of the Recovery Act award was to provide funds to hire a new technician to expedite completion of the parent project. Specifically, this new hire will be responsible for the following experiments: 1) Optimization of conditions to achieve efficient RNAi-based knockdown of cellular enzymes with key roles in mRNA degradation, 2) Preparation of cytoplasmic extracts and purification of recombinant proteins necessary to develop an in vitro assay for SOX activity, and 3) Affinity purification of antibodies for protein interaction experiments, as well as optimization of conditions for such experiments. I have hired the technician, and his training period is now underway He is currently in the process of generating and purifying recombinant proteins for the establishment of in the in vitro assay. He is also participating in production and purification of polyclonal antibodies for protein interaction experiments. It should be noted that because funds were deposited in my account on September 15, payroll for the technician will not be posted until the first week of October (after the current reporting deadline).
Jobs Summary: A technician was hired to assist directly with this project. (Total jobs reported: 0)
Project Status: Less Than 50% Completed
This award's data was last updated on Sep. 15, 2009. Help expand these official descriptions using the wiki below.
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