RUI - Functional Characterization of a Nematode Gene Related to the Class II bHLH Protein, ABF-1

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Basic helix-loop-helix proteins are important transcriptional regulators of cellular proliferation and differentiation in organisms ranging from invertebrates to vertebrates. Recently, a new gene closely related to the mammalian basic helix-loop-helix protein activated B-cell factor 1 (ABF-1), designated CeABF-1, was cloned from the nematode Caenorhabditis elegans. The objectives of the research include 1) investigating the function of the CeABF-1 gene; 2) elucidating the transcriptional properties of the CeABF-1 protein; 3) identifying the target gene(s) regulated by CeABF-1. To accomplish these specific goals, gene function and expression studies will be performed to delineate the role of CeABF-1 in the nematode. The functional studies will integrate a broad spectrum of genetic and biochemical techniques, including RNA microinjections and the development of transgenic animals for expression analyses. The transcriptional properties and target gene(s) regulated by CeABF-1 will be analyzed using DNA transfections, electrophoretic-mobility shift assays and DNA microanalysis. This project will allow for the comparison of HLH gene function in both nematodes and humans and identify conserved molecular pathways important for cellular processes. The results of these studies will help to elucidate the evolutionary as well as the biological relationship between the nematode and human ABF-1 protein. Helix-loop-helix (HLH) proteins are important transcriptional regulators of cellular proliferation and differentiation in organisms ranging from invertebrates to vertebrates. The importance of HLH proteins as potent mediators of immune system function and muscle development has been well established. The significance of these studies will allow for the determination of the biological function of the nematode Caenorhabditis elegans HLH gene, designated CeABF-1, and will provide data that will help to elucidate the molecular mechanisms that govern cell-restricted eukaryotic gene expression. This project will enhance the knowledge of target gene(s) controlled by the protein CeABF-1, and provide an understanding of the ancestral relationship between the worm CeABF-1 and human homologue, activated B-cell factor 1 (ABF-1).