NSF Award
9418632
9418632 Perry The overall goal of this research is to elucidate molecular mechanisms that govern gene expression in eukaryotic cells. The proposed studies are aimed at understanding the role of an interesting DNA-binding protein, CHD1, in the determination of higher-order chromatin structure and the regulation of gene activity. CHD1 possesses certain structural features (a chromodomain and an ATPase/helicase domain), which relate it to other groups of chromatin-associated proteins that have been implicated in transcriptional regulation. A broad spectrum of genetic and biochemical techniques will be used, including DNA transfection and microinjection experiments in a variety of cultured cells, immunofluorescence analyses, protein purification by column chromatography, gene cloning and sequencing, DNA-, RNA- and immuno-blotting, primer extension and nucleaseprotection assays, electrophoretic-mobility shift and DNA footprint analyses, and the production of wild-type and mutant proteins in bacteria. The studies will be carried out with mammalian cells, Drosophila and yeast. The results of these studies should contribute substantially to our knowledge of gene regulation by defining factors that determine the accessibility of genes to the transcriptional apparatus and that help mediate cell-cycle dependent changes in chromatin condensation. Although an impressive body of facts pertaining to gene regulation has accumulated over the past several years, the general problems being addressed here remain largely unsolved. %%% The overall goal of this research is to elucidate molecular mechanisms that govern gene expression in eukaryotic cells. The proposed studies are aimed at understanding the role of an interesting DNA-binding protein, CHD1, in the determination of higher-order chromatin structure and the regulation of gene activity, CHD1 possesses certain structural features which relate it to other groups of chromatin-associated proteins that have been implicated in transcriptional regulation. A broad spectrum of genetic and biochemical investigative techniques will be used and studies will be carried out with mammalian cells, Drosophila and yeast. Although an impressive body of facts pertaining to gene regulation has accumulated over the past several years, the general problems being addressed here remain largely unsolved. The results of these studies should contribute substantially to our knowledge of gene regulation by defining factors that determine the accessibility of genes to the transcriptional apparatus and that help mediate cell-cycle dependent changes in chromatin structure. *** ??
