MiCLIP-MaPseq, a Substrate Identification Approach for Radical SAM RNA Methylating Enzymes.

ABSTRACT: Although present across bacteria, the large family of radical SAM RNA methylating enzymes is largely uncharacterized. Escherichia coli RlmN, the founding member of the family, methylates an adenosine in 23S rRNA and several tRNAs to yield 2-methyladenosine (m²A). However, varied RNA substrate specificity among RlmN enzymes, combined with the ability of certain family members to generate 8-methyladenosine (m⁸A), makes functional predictions across this family challenging. Here, we present a method for unbiased substrate identification that exploits highly efficient, mechanism-based cross-linking between the enzyme and its RNA substrates. Additionally, by determining that the thermostable group II intron reverse transcriptase introduces mismatches at the site of the cro

SUBMITTER: Stojkovic V 

PROVIDER: S-EPMC7207077 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature