Defect in the Formation of 70S Ribosomes Caused by Lack of Ribosomal Protein L34 Can Be Suppressed by Magnesium

• DOI: 10.1128/jb.01896-14
• Publication Date: 2014-09-03T12:10:53Z
• Authors: Genki Akanuma; Ako Kobayashi; Shota Suzuki; Fujio Kawamura; Yuh Shiwa; Satoru Watanabe; Hirofumi Yoshikawa; Ryo Hanai; Morio Ishizuka
• Keywords: Ribosomal Proteins; Time Factors; Bacterial Proteins; Genetic Complementation Test; Mutation; Magnesium; Gene Expression Regulation, Bacterial; Ribosomes; Bacillus subtilis

ABSTRACT

To elucidate the biological functions of the ribosomal protein L34, which is encoded by the rpmH gene, the rpmH deletion mutant of Bacillus subtilis and two suppressor mutants were characterized. Although the Δ rpmH mutant exhibited a severe slow-growth phenotype, additional mutations in the yhdP or mgtE gene restored the growth rate of the Δ rpmH strain. Either the disruption of yhdP , which is thought to be involved in the efflux of Mg 2+ , or overexpression of mgtE , which plays a major role in the import of Mg 2+ , could suppress defects in both the formation of the 70S ribosome and growth caused by the absence of L34. Interestingly, the Mg 2+ content was lower in the Δ rpmH cells than in the wild type, and the Mg 2+ content in the Δ rpmH cells was restored by either the disruption of yhdP or overexpression of mgtE. In vitro experiments on subunit association demonstrated that 50S subunits that lacked L34 could form 70S ribosomes only at a high concentration of Mg 2+ . These results showed that L34 is required for efficient 70S ribosome formation and that L34 function can be restored partially by Mg 2+ . In addition, the Mg 2+ content was consistently lower in mutants that contained significantly reduced amounts of the 70S ribosome, such as the Δ rplA (L1) and Δ rplW (L23) strains and mutant strains with a reduced number of copies of the rrn operon. Thus, the results indicated that the cellular Mg 2+ content is influenced by the amount of 70S ribosomes.