TERWILLIGER, N.B.**; RYAN , M.; PHILLIPS, M.; University of Oregon; University of Oregon; University of Oregon: Crustacean Hemocyanin Gene Family: Oxygen Probes of the Global Gene Scene

Global expression profiling simultaneously monitors vast numbers of genes in an organism, and it identifies groups of co-regulated genes. The principles of molecular recognition at the genomic and proteomic levels were developed for single gene analysis, and data analysis of global expression profiling requires knowledge of gene families and products. Oxygen delivery in decapod crustaceans is provided by hemocyanin (Hc). The Hc gene family, like hemoglobin, has undergone a process of gene duplication and divergence. These events have resulted in a Hc molecule composed of multiple subunits with a functional spectrum of homeotropic and heterotropic responses to allosteric effectors, metabolites and environmental factors. Other duplication events have resulted in new proteins with marked changes in function, as evidenced by phenoloxidase, cryptocyanin, and insect hexamerins. Continuing investigations at the gene family level in concert with genomics and proteomics will provide new insights into crustacean physiology. We have determined the cDNA sequences of all six subunits that compose the Hc molecule in the adult Dungeness crab, Cancer magister. The subunits are the products of six separate genes. Analyses of protein and gene structures of these Hc subunits confirm the clustering into two motifs. Hc genes 1 and 2 are tandemly linked on the same chromosome and have one intron. Genes 3,4,5 and 6 have an additional intron. The exon-intron pattern in C. magister will facilitate Hc gene structure comparisons in the Arthropoda and other phyla. Analyses of upstream regulatory regions of the Hc genes in conjunction with ongoing development and construction of a C. magister microarray will yield insights into transcriptome changes during development, molting and environmental stress. Supported by NSF 9984202.