2015 Fiscal Year Annual Research Report
Role of RNA methylation in the regulation of gene expression.
| Project/Area Number |
26870283
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| Research Institution | Kyoto University |
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Principal Investigator |
FUSTIN JM 京都大学, 薬学研究科(研究院), 講師 (50711818)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Keywords | m6A methylation / phosphoproteomics |
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| Outline of Annual Research Achievements |
Our investigations have proceeded well over the last year since we are currently writing our next manuscript reporting our latest investigations.
We have quantified m6A RNA methylation of the liver transcriptome every 4 hours during 24 hours and have identified significant methylation sites. Most of these methylation sites were detected at all time points but some methylation sites were only detected at specific times.We have then focussed on methylations sites detected at all time points and identified that one candidate transcript in particular, with a very important role in the circadian clock, is highly methylated. By using a methylation inhibitor and knocking down the methylransferases Mettl3 and Mettl14, we have observed that the expression of the protein encoded by this transcript increases. Interestingly, we have identified two alternative transcripts that are both affected by RNA methylation inhibition but in different ways, leading to an increase in their encoded proteins.
Next, we have investigated the function of these two isoforms in the circadian clock and observed that they have opposite functions: one isoform causes period elongation, the other period shortening. Using protein level analyses and phosphoproteomics we have then precisely identified the roles of these two isoforms.
As previously mentioned we are currently in the process of writing a manuscript reporting these observations.
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