| Budget Amount *help |
¥3,420,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥420,000)
Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2010: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
In order to utilize cryptochrome(CRY) as a molecular light switch for regulation of gene activity, we characterize CRYs in a variety of animals. Complementary DNA encoding cryptochromes were isolated from vertebrates such as Xenopus, quail, goldlined spinefoot. Because these CRYs are considered to interact with circadian clock components such as CLOCK and BMALs, we investigated whether the CRY proteins could interact with and inhibit the activities of those clock proteins by transcriptional analysis. Xenopus CRYs(XtCRY1 and XtCRY2) inhibited the transcriptional activities, suggesting that those CRYs work as circadian clock components in Xenopus. Next, we investigated mRNA expression of CRYs in the goldlined spinefoot, and observed that the CRY mRNA levels showed clear lunar rhythms with their peak at the first quarter moon. This strongly suggests that CRYs function for not only circadian clock but also lunar clock.
Next, we searched putative CRY-interacting proteins by yeast two-hybrid screening. We identified several possible interactors, and interestingly, some of them showed light-dependent interaction, showing that they can be used as light-dependent switchable molecules. This system can be applied to regulation of gene activity by light at the transcription levels, and future experiments should be planned for light-dependent regulation of the protein localization, interaction, etc.
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