2014 Fiscal Year Final Research Report
Base excising restriction enzymes
| Project/Area Number |
26650123
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Genetics/Chromosome dynamics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
YANO Hirokazu 東京大学, 大学院新領域創成科学研究科, 特任助教 (20646773)
IDE Hiroshi 広島大学, 理学(系)研究科(研究院), 教授 (30223126)
FUKUYO Masaki 千葉大学, 医学研究院, 特任助教 (40639085)
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| Project Period (FY) |
2014-04-01 – 2015-03-31
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| Keywords | エピジェネティクス / 制限酵素 / 制限修飾系 / 細菌 / 塩基除去 / AP リアーゼ / DNA グリコシラーゼ |
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| Outline of Final Research Achievements |
So far, all the restriction enzymes hydrolyze phosphodiester bonds linking the monomer units of DNA. We recently reported that a restriction enzyme (R.PabI) of the PabI superfamily with half-pipe fold has DNA glycosylase activity that excises an adenine base in the recognition sequence (5′-GTAC). We now found a second activity in this enzyme: at the resulting apurinic/apyrimidinic (AP) (abasic) site (5′-GT#C, # = AP), its AP lyase activity generates an atypical strand break. Its covalent DNA-R.PabI reaction intermediates can be trapped by NaBH4 reduction. The base excision is not coupled with the strand breakage and yet causes restriction because the restriction enzyme action can impair transformation ability of unmethylated DNA even in the absence of strand breaks in vitro. The base excision of R.PabI is inhibited by methylation of the target adenine base. These findings expand our understanding of genetic and epigenetic processes linking those in prokaryotes and eukaryotes.
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| Free Research Field |
ゲノム動態学
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