I trained in Internal Medicine at the University of California at San Francisco and have fellowship training in Cardiology from Duke University and Infectious Diseases from the University of Washington, Seattle. My immediate goal is to complete my scientific training so that I will possess the necessary tools to pursue my long term objective of becoming an independent physician-researcher involved in the investigation of the relationship between viral infection and diseases of the cardiovascular system at the molecular level. This goal will be realized by means of an intensive engagement in focused coursework and basic research to be carried out at the Fred Hutchinson Cancer Research Center and the University of Washington, Seattle over a period of five years. The long term goals of this project are to understand the process of translational regulation as it is mediated by upstream open reading frames (uORFs) in Human Cytomegalovirus(CMV). CMV is an important pathogen which causes disease in immunocompromised patients and neonates. There is mounting evidence that CMV may play a role in the process of coronary atherosclerosis. We believe that translational regulation is an important means of controlling gene expression in CMV. The process of translational regulation may be an important mechanism of gene regulation in mammalian systems as well. Approximately two-thirds of oncogenes, growth factors, and growth factor receptors have uORFs in the 5' leader sequences of their mRNAs, yet our understanding of their purpose and mechanism of action is primitive. This proposed research seeks to learn more about translational regulation by studying the CMV gene UL5. There is evidence to suggest that this gene is expressed and is translated from a polycistronic message. The 5' leader sequence of UL5 contains no less than twelve AUG codons with associated uORFs. UL5 may be a promising model for the study of translational regulation.
The specific aims of the research are l) to determine whether UL5 is expressed and 2) to discover the mechanism by which it is translationally regulated.
Specific aim #1 will be accomplished by generating polyclonal antisera against UL5 and using it to identify, locate, and characterize the protein and its expression. We will synthesize UL5 in a bacterial expression system as a histidine-tagged fusion protein and isolate it by means of affinity chromatography.
Specific aim #2 will be accomplished using a mutational analysis of the 5' leader of UL5 to assess its influence on the translational activity of UL5. A UL5/B-gal fusion gene will serve as a reporter.
