Methyltransferase that modifies guanine 966 of the 16 S rRNA: Functional identification and tertiary structure

Dmitry V. Lesnyak, Jerzy Osipiuk, Tatiana Skarina, Petr V. Sergiev, Alexey A. Bogdanov, Aled Edwards, Alexei Savchenko, Andrzej Joachimiak, Olga A. Dontsova

Research output: Contribution to journalArticlepeer-review

69 Citations (Scopus)

Abstract

N2-Methylguanine 966 is located in the loop of Escherichia coli 16 S rRNA helix 31, forming a part of the P-site tRNA-binding pocket. We found yhhF to be a gene encoding for m2G966 specific 16 S rRNA methyltransferase. Disruption of the yhhF gene by kanamycin resistance marker leads to a loss of modification at G966. The modification could be rescued by expression of recombinant protein from the plasmid carrying the yhhF gene. Moreover, purified m2G966 methyltransferase, in the presence of S-adenosylomethionine (AdoMet), is able to methylate 30 S ribosomal subunits that were purified from yhhF knock-out strain in vitro. The methylation is specific for G966 base of the 16 S rRNA. The m2G966 methyltransferase was crystallized, and its structure has been determined and refined to 2.05 Å. The structure closely resembles RsmC rRNA methyltransferase, specific for m2G1207 of the 16 S rRNA. Structural comparisons and analysis of the enzyme active site suggest modes for binding AdoMet and rRNA to m 2G966 methyltransferase. Based on the experimental data and current nomenclature the protein expressed from the yhhF gene was renamed to RsmD. A model for interaction of RsmD with ribosome has been proposed.

Original languageEnglish
Pages (from-to)5880-5887
Number of pages8
JournalJournal of Biological Chemistry
Volume282
Issue number8
DOIs
Publication statusPublished - 23 Feb 2007
Externally publishedYes

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