COHEN*, AARON/J*; POPE, ROBERT/K; Indiana University South Bend; Indiana University South Bend: Cloning and Analysis of Zebrafish Supervillin

Supervillin is a 205-kDa F-actin binding protein originally identified in bovine neutrophils. Subsequently, supervillin has been identified in human and murine tissues. Distribution of mRNA for supervillin has shown that this protein is expressed in most body tissues, with the exception of nervous tissue. Recently, a muscle specific 250-kDa splice variant (archvillin) has been identified from human skeletal muscle. Examination of these cDNA sequences shows that supervillin/archvillin are bipartite proteins, containing a highly conserved carboxy-terminal half that is very homologous to villin, and an amino-terminal half that is entirely novel. The amino acid sequence of this protein has numerous sequences for nuclear localization, sites for serine/threonine phosphorylation, a site for tyrosine phosphorylation, and numerous mapped and unmapped F-actin binding sites. Previous research has demonstrated localization of supervillin at the plasma membrane of cells that are confluent, and nuclear localization in cells that are subconfluent. In this research we have cloned and sequenced the full-length zebrafish supervillin cDNA. The ability to examine the function of this protein in zebrafish will enable us to elucidate the possible roles of supervillin in the cell. The zebrafish model system will enable us to utilize biochemistry, cell biology, and molecular biology techniques to identify the expression/role of this protein during embryonic development, tissue differentiation, and to identify the function of this protein in adult specimens.