Requlation of gene expression in sea urchin embryo

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This research project has been proposed to obtain clues to understand a mechanism underlying temporal and spatial expression of genes during early development of sea urchin. For this purpose we selected a gene for arylsulfatase (Ars) that expresses in the temporally and spatially regulated manner in sea urchin embryo. The sea urchin Ars gene contains a TATA box in its basal promoter, several G-string sequences and a long polypyrimidine region in its 5' flanking region. Following results were obtained in this study. (1) The long polypyrimidine region can form a triplex DNA configurationthrough Hoogsten base parings, (2) four cDNA clones for G-string DNA-binding proteins were obtained by south-western blotting using a synthetic poly (dG)-poly(dC) as the probe. All these cDNAs contain GA-rich sequence in its ORF.(3) Temporal expression of the Ars gene is primarily determined by a short region between -1 and -100b positions. Two cis-elements between -100 and -252 position seem to be related to temporal regulation of the Ars gene expression. (4) A 229nts sequence in the first intron strongly enhances the activity of these elements. all these results were obtained by luciferase assays. (5) A nuclear extract system to effectively transcribe the Ars gene in vitro was constructed.