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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
Formation and removal of 1,N6-dimethyladenosine in mammalian transfer RNA
Xue-Jiao You1,2,3, Shan Zhang1,2, Juan-Juan Chen2
1Department of Radiation and Medical Oncology, Cancer Precision Diagnosis and Treatment and Translational Medicine Hubei Engineering Research Center, Zhongnan Hospital of Wuhan University, School of Public Health, Wuhan University, Wuhan 430071, China.
Abstract:
RNA molecules harbor diverse modifications that play important regulatory roles in a variety of biological processes. Over 150 modifications have been identified in RNA molecules. N6-methyladenosine (m6A) and 1-methyladenosine (m1A) are prevalent modifications occurring in various RNA species of mammals. Apart from the single methylation of adenosine (m6A and m1A), dual methylation modification occurring in the nucleobase of adenosine, such as N6,N6-dimethyladenosine (m6,6A), also has been reported to be present in RNA of mammals. Whether there are other forms of dual methylation modification occurring in the nucleobase of adenosine other than m6,6A remains elusive. Here, we reported the existence of a novel adenosine dual methylation modification, i.e. 1,N6-dimethyladenosine (m1,6A), in tRNAs of living organisms. We confirmed that m1,6A is located at position 58 of tRNAs and is prevalent in mammalian cells and tissues. The measured level of m1,6A ranged from 0.0049% to 0.047% in tRNAs. Furthermore, we demonstrated that TRMT6/61A could catalyze the formation of m1,6A in tRNAs and m1,6A could be demethylated by ALKBH3. Collectively, the discovery of m1,6A expands the diversity of RNA modifications and may elicit a new tRNA modification-mediated gene regulation pathway.
Insights
Researchers discovered a new dual methylation modification in RNA, 1,N6-dimethyladenosine (m1,6A), found in transfer RNAs (tRNAs) of mammals. This finding expands the known landscape of RNA modifications and suggests new regulatory pathways.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- RNA molecules contain over 150 diverse modifications crucial for biological regulation.
- N6-methyladenosine (m6A) and 1-methyladenosine (m1A) are common adenosine modifications in mammals.
- Dual methylation modifications like N6,N6-dimethyladenosine (m6,6A) have been identified, but others remain unknown.
Purpose of the Study:
- To identify novel dual methylation modifications in adenosine.
- To investigate the presence and characteristics of 1,N6-dimethyladenosine (m1,6A) in RNA.
Main Methods:
- Analysis of RNA modifications in mammalian cells and tissues.
- Mass spectrometry or similar techniques to identify and quantify m1,6A.
- Enzymatic assays to determine the synthesis and demethylation pathways of m1,6A.
Main Results:
- A novel adenosine dual methylation modification, 1,N6-dimethyladenosine (m1,6A), was discovered in transfer RNAs (tRNAs).
- m1,6A was confirmed to be located at position 58 of tRNAs and is prevalent in mammalian cells.
- The levels of m1,6A in tRNAs were quantified, ranging from 0.0049% to 0.047%.
- TRMT6/61A was identified as the enzyme catalyzing m1,6A formation, and ALKBH3 as the demethylating enzyme.
Conclusions:
- The discovery of m1,6A expands the known diversity of RNA modifications.
- m1,6A is a prevalent tRNA modification in mammals.
- This finding may reveal new tRNA modification-mediated gene regulation pathways.
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