A chemical screen for biological small molecule-RNA conjugates reveals CoA-linked RNA.

Proc Natl Acad Sci U S A
Authors
Keywords
Abstract

Compared with the rapidly expanding set of known biological roles for RNA, the known chemical diversity of cellular RNA has remained limited primarily to canonical RNA, 3'-aminoacylated tRNAs, nucleobase-modified RNAs, and 5'-capped mRNAs in eukaryotes. We developed two methods to detect in a broad manner chemically labile cellular small molecule-RNA conjugates. The methods were validated by the detection of known tRNA and rRNA modifications. The first method analyzes small molecules cleaved from RNA by base or nucleophile treatment. Application to Escherichia coli and Streptomyces venezuelae RNA revealed an RNA-linked hydroxyfuranone or succinyl ester group, in addition to a number of other putative small molecule-RNA conjugates not previously reported. The second method analyzes nuclease-generated mononucleotides before and after treatment with base or nucleophile and also revealed a number of new putative small molecule-RNA conjugates, including 3'-dephospho-CoA and its succinyl-, acetyl-, and methylmalonyl-thioester derivatives. Subsequent experiments established that these CoA species are attached to E. coli and S. venezuelae RNA at the 5' terminus. CoA-linked RNA cannot be generated through aberrant transcriptional initiation by E. coli RNA polymerase in vitro, and CoA-linked RNA in E. coli is only found among smaller (approximately

Year of Publication
2009
Journal
Proc Natl Acad Sci U S A
Volume
106
Issue
19
Pages
7768-73
Date Published
2009 May 12
ISSN
1091-6490
DOI
10.1073/pnas.0900528106
PubMed ID
19416889
PubMed Central ID
PMC2674394
Links
Grant list
R01 GM065865 / GM / NIGMS NIH HHS / United States
R01GM065865 / GM / NIGMS NIH HHS / United States
Howard Hughes Medical Institute / United States