A large body of evidence supports the notion that Myc plays a pivotal role
in carcinogenesis. Myc has been shown to function as a transcription
factor, yet the critical target genes that it regulates and the manner in
which these impact on cell behavior have yet to be rigorously delineated.
Quantitative proteome analysis is the global analysis of protein expression and has a number of advantages over mRNA expression analysis. It directly focuses on the actual biological effector molecules and provides more accurate information about biological systems. The recently developed ICAT (isotope coded affinity tags) technology greatly expands the range of proteins that can be analyzed and allows the accurate quantitation and concurrent sequence identification of the individual proteins in complex mixtures.
This study aims at the application of quantitative proteomics to the analysis
of the function of the Myc oncoprotein. Specifically, ICAT technology
will be used to identify global and location-specific protein changes induced
by Myc. The status of chromatin-associated regulatory factors in Myc-OFF
and Myc-ON B lymphocytes or in growing fibroblasts and fibroblasts arrested
by Mad (Myc antagonist) will be compared by the combination of chromatin
isolation and ICAT analysis. ICAT technology will also be used to analyze
the protein changes during induction of apoptosis by Myc.
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