| Budget Amount *help |
¥40,430,000 (Direct Cost: ¥31,100,000、Indirect Cost: ¥9,330,000)
Fiscal Year 2016: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Fiscal Year 2015: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000)
Fiscal Year 2014: ¥12,740,000 (Direct Cost: ¥9,800,000、Indirect Cost: ¥2,940,000)
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| Outline of Final Research Achievements |
Rif1 is a conserved chromatin factor that regulates DNA replication, DNA repair and transcription. It may be involved in chromatin loop formation at nuclear periphery. We have determined genome-wide Rif1 bindings sites (Rif1BS) in fission yeast and identified a G-rich consensus sequence (Rif1CS). We then demonstrated the essential role of Rif1CS in chromatin binding of Rif1 and subsequent repression of DNA replication over a 100kb segment. Footprinting with DMS or various nucleases, polymerase stop assays as well as the effect of 7-deaza dGTP demonstrated that Rif1BS adopts G-quadruplex like structures. Purified fission yeast and mammalian Rif1 proteins specifically bind to this structure. Rif1 can simultaneously bind to multiple molecules of G4 through oligomerization, providing potential basis for its ability to tether multiple chromatin fibers to generate chromatin compartment that may coordinately regulate replication, transcription or repair.
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