International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


POSTER 131 - FUNCTIONAL ANALYSIS OF CREB BINDING PROTEIN (CBP) IN MICE USING AN ENU-BASED MUTAGENESIS APPROACH IN EMBRYONIC STEM CELLS

M.K. Bunger
University of North Carolina

Magnuson TR
University of North Carolina

The CREB binding protein (CBP) is a transcriptional coactivator with histone acetyltransferase (HAT) activity.  Through this activity, CBP integrates multiple biological signals that are critical for developmental and physiological processes. Several gene knockout studies indicate that functional CBP protein is necessary for embryonic development in mice. Moreover, heterozygotes of these null alleles also exhibit phenotypes resembling the human disease Rubenstein-Taybi syndrome characterized by specific limb defects,  lymphoid tumors, and learning deficits. We are using a novel gene-based mutagenesis strategy in embryonic stem cells to generate an allelic series of mutations at Cbp in mice in order to address specific roles of subdomains of CBP protein in vivo.  Several point mutations identified alter important amino acids and splicing in domains of CBP protein responsible for HAT activity and protein-protein interactions.  Once mice are made, a battery of in-vitro, tumor, vascular, behavioral, and skeletal endpoints will be assessed to provide insight into specific contributions of individual CBP domains and interactions in mouse development and disease processes associated with loss of CBP.  This mutagenesis strategy allows us to know the nature of each amino acid substitution before animals are made. Therefore will also discuss in silico approaches being undertaken to help make functional predictions of our identified amino acid substitutions in efforts to streamline mouse production


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