| 研究実績の概要 |
Sialic acid (Sia) has a variety of natural modifications, which are often regulated developmentally and organ-specificically. However, the significance of the structural diversity and the spatiotemporally regulated expression of Sia have remained largely mysterious up to now. Thus, our long goal is to give an insight into biological function and its molecular mechanisms of Sia. For this purpose, we focus on CMP-Sia synthetase (CSS): (1) To reveal the significance of the structural diversity and the spatiotemporally regulated expression of Sia-glycoconjugates at an animal level; (2) To understand how important CSS is in invertebrates, focusing on the intracellular localization and the role of C-terminal half domain.
For Objective (1), CSS knock-down (KD) medakas were generated using TILLING methods. Two mutant medaka strains I230N and L303Q, which had very low in vitro activity, were established. CSS knock-out (KO) medaka was also made by TALEN methods.
For Objective-(2), the CSS cDNAs were cloned from Tribolium castaneum (red flour beetle), Aedes aegypti (yellow fever mosquito), and Drosophila melanogaster (fruit fly) to analyze kinetics for Sia species and intracellular localization using the recombinant enzymes.
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