| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Outline of Final Research Achievements |
This project aimed to understand a relationship of lysine (Lys) acetylation and translational regulation in bacteria. I focused on the acetylation introduced in Lys-74 position of alanyl-tRNA synthetase (AlaRS) in E. coli because the Lys is essential for the enzymatic activity. I incorporated an acetyllysine at the position by genetic code expansion technology with AcKRS/tRNA(Pyl) pair. The activity of the mutant, AlaRS K74Ac was affected by the acetylation, high level of which was strongly impaired the activity in vitro. Although the acetylation was eliminated by deacetylase (CobB) in vivo, the deactylation was a little in vitro. Based on the previous report that CobB interacts with ribosome, I hypothesized that this deacetylation might occur on ribosome. I constructed a strain expressing CobB mutant which does not interact with ribosome. Since the AlaRS K74Ac prepared from the strain was also deacetylated in vivo, it was suggested a different mechanism from my idea exists in E. coli.
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