Control Of Mammalian Cell Proliferation

Child Health And Human Development

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Abstract

The long term goals of the Human Cell Genetics Section (HCGS) are: i) to elucidate mechanisms by which determinants of higher order chromatin structure and associated epigenetic mechanisms regulate cell growth and survival; and ii) to relate such cellular control mechanisms to senescence and age-related disease in the intact organism. Of particular interest is the hypothesis that damage and remodeling of chromatin-based structures contribute to mammalian senescence, both at the cellular and organismal levels. One approach to detect such remodeling is a semi-random sampling of genome chromatin structure, DDChIP, a method similar in principle to differential display. Over the past year, a second approach has been added. The latter is based on bioinformatics software developed by the HCGS. Sequence motifs or clusters of motifs are searched for in the human and mouse genomes. Further criteria to define objects of interest include inverted repeats, nearby CpG islands, etc. Loci identified in the two genomes are assessed for conservation by cross-blast analysis. This step eliminates the vast majority of noise in the search results. Lists of candidate loci are submitted for automated primer design and synthesis. Normalized arrays of primer pairs facilitate robot-based PCR assembly. Finally, a novel automated HPLC-based technique for PCR analysis coupled with custom graphics software allows rapid evaluation of the results. Current experiments are exploring the utility of this bioinformatics-experiment interface in pursuing the above stated goals.