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PMID: 18668122 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The RNA acetyltransferase driven by ATP hydrolysis synthesizes N4-acetylcytidine of tRNA anticodon.

The EMBO journal ·Vol. 27 ·No. 16 ·2008-08-20 ·Pages 2194-203

Ikeuchi Y, Kitahara K, Suzuki T

Abstract

The wobble base of Escherichia coli elongator tRNA(Met) is modified to N(4)-acetylcytidine (ac(4)C), which is thought to ensure the precise recognition of the AUG codon by preventing misreading of near-cognate AUA codon. By employing genome-wide screen of uncharacterized genes in Escherichia coli ('ribonucleome analysis'), we found the ypfI gene, which we named tmcA (tRNA(Met) cytidine acetyltransferase), to be responsible for ac(4)C formation. TmcA is an enzyme that contains a Walker-type ATPase domain in its N-terminal region and an N-acetyltransferase domain in its C-terminal region. Recombinant TmcA specifically acetylated the wobble base of E. coli elongator tRNA(Met) by utilizing acetyl-coenzyme A (CoA) and ATP (or GTP). ATP/GTP hydrolysis by TmcA is stimulated in the presence of acetyl-CoA and tRNA(Met). A mutation study revealed that E. coli TmcA strictly discriminates elongator tRNA(Met) from the structurally similar tRNA(Ile) by mainly recognizing the C27-G43 pair in the anticodon stem. Our findings reveal an elaborate mechanism embedded in tRNA(Met) and tRNA(Ile) for the accurate decoding of AUA/AUG codons on the basis of the recognition of wobble bases by the respective RNA-modifying enzymes.

MeSH Terms
Acetyltransferases/chemistry,metabolism Adenosine Triphosphate/metabolism Amino Acid Sequence Anticodon/metabolism Base Sequence Cytidine/analogs & derivatives,biosynthesis,chemistry Escherichia coli/enzymology,genetics Escherichia coli Proteins/chemistry,metabolism Genes, Bacterial Guanosine Triphosphate/metabolism Hydrolysis Kinetics Mass Spectrometry Molecular Sequence Data Mutation/genetics Nucleic Acid Conformation RNA, Transfer, Met/chemistry,genetics,metabolism Recombinant Proteins Ribosomes/metabolism Sequence Alignment
Chemicals
Anticodon Escherichia coli Proteins RNA, Transfer, Met Recombinant Proteins N-acetylcytidine Cytidine Guanosine Triphosphate Adenosine Triphosphate Acetyltransferases TmcA protein, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ikeuchi Yoshiho
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Kitahara Kei
Suzuki Tsutomu
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2008.154
Published
2008-08-20
Epub
2008-00-31
Pages
2194-203
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2500205
Subset
IM
External Links
PubMed source DOI: 10.1038/emboj.2008.154 PMC Full Text View journal details
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