Epitranscriptome analysis of sporulating budding yeast

2014 Fiscal Year Final Research Report

• Project/Area Number: 25620125
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: SUZUKI Tsutomu 東京大学, 工学(系)研究科(研究院), 教授 (20292782)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: RNA修飾 / N6メチルアデノシン

Outline of Final Research Achievements

Eukaryotic mRNAs and non-coding RNAs contain various post-transcriptional modifications including 5’ cap structure, inosine (I), 5-methylcytidine (m5C), and N6-methyladenosine (m6A). To investigate functional roles of m6A, we screened m6A sites on mRNAs in sporulating budding yeast. More than 8,000 candidate regions were mapped in about 3,000 genes in Saccharomyces cerevisiae by m6A-seq which is a biochemical method to identify m6A regions by immunoprecipitation of m6A-containing RNA fragment by anti-m6A antibody combined with deep sequencing. In addition, we identified a single m6A site in IME1 mRNA by RNA mass spectrometric analysis. However, this m6A site was not confirmed by m6A-seq, exposing a problem of anti-m6A antibody with poor specificity.

Free Research Field

分子生物学