YhiQ Is RsmJ, the Methyltransferase Responsible for Methylation of G1516 in 16S rRNA of E. coli

• Published in: Journal of molecular biology 2012-01, Vol.415 (1), p.16-21
• Main Authors: Basturea, Georgeta N., Dague, Darryl R., Deutscher, Murray P., Rudd, Kenneth E.
• Format: Article
• Language: English
• Subjects: Base Sequence; E. coli; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; genes; genetics; metabolism; Methylation; methyltransferase; methyltransferases; Methyltransferases - genetics; Methyltransferases - metabolism; Models, Molecular; Molecular Sequence Data; Nucleic Acid Conformation; Phenotype; reverse transcription; ribosomal RNA; Ribosomes; Ribosomes - genetics; Ribosomes - metabolism; RNA modification; RNA, Ribosomal, 16S; RNA, Ribosomal, 16S - genetics; RNA, Ribosomal, 16S - metabolism; small ribosomal subunit; Substrate Specificity
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2011.10.044

Ten methyltransferases and one pseudouridine synthase are required for complete modification of the small ribosomal subunit in Escherichia coli. Nine methyltransferases, as well as the pseudouridine synthase, are already known. Here, we identify RsmJ, the last unknown methyltransferase required for methylation of m2G1516 in 16S ribosomal RNA (rRNA), as the protein encoded by yhiQ. Reverse transcription primer extension analysis reveals that rRNA extracted from a yhiQ deletion strain is not methylated at G1516. Moreover, methylation is restored upon gene complementation. Also, purified recombinant YhiQ specifically methylates 30S subunits extracted from the deletion strain. The absence of the yhiQ gene leads to a cold-sensitive phenotype. Based on these data, we propose that the yhiQ gene be renamed rsmJ. [Display omitted] ► A yhiQ deletion mutant is missing the 16S rRNA m2G1516 modification. ► Purified YhiQ methylates isolated yhiQ mutant 30S ribosomal subunits in vitro. ► A yhiQ mutant strain exhibits a cold-sensitive growth defect. ► YhiQ is identified as the RsmJ 16S G1516 rRNA methyltransferase.