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Journal of Bacteriology, July 2009, p. 4555-4561, Vol. 191, No. 14
0021-9193/09/$08.00+0     doi:10.1128/JB.00263-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Transcription Activity of Individual rrn Operons in Bacillus subtilis Mutants Deficient in (p)ppGpp Synthetase Genes, relA, yjbM, and ywaC{triangledown} ,{dagger}

Yousuke Natori,1,{ddagger} Kazumi Tagami,1,{ddagger} Kana Murakami,1 Sawako Yoshida,1 Osamu Tanigawa,1 Yoonsuh Moh,1 Kenta Masuda,1 Tetsuya Wada,1 Shota Suzuki,1 Hideaki Nanamiya,2 Yuzuru Tozawa,2 and Fujio Kawamura1*

Laboratory of Molecular Genetics and Research Information Center for Extremophile, College of Science, Rikkyo University, Toshima-ku Nishi-ikebukuro 3-34-1, Tokyo 171-8501, Japan,1 Cell-free Science and Technology Research Center, Ehime University, Bunkyo-cho, Matsuyama 790-8577, Japan2

Received 27 February 2009/ Accepted 2 May 2009

In Bacillus subtilis a null mutation of the relA gene, whose gene product is involved in the synthesis and/or hydrolysis of (p)ppGpp, causes a growth defect that can be suppressed by mutation(s) of yjbM and/or ywaC coding for small (p)ppGpp synthetases. All 35 suppressor mutations newly isolated were classified into two groups, either yjbM or ywaC, by mapping and sequencing their mutations, suggesting that there are no (p)ppGpp synthetases other than RelA, YjbM, and YwaC in B. subtilis. In order to understand better the relation between RelA and rRNA synthesis, we studied in the relA mutant the transcriptional regulation of seven rRNA operons (rrnO, -A, -J, -I, -E, -D, or -B) individually after integration of a promoter- and terminatorless cat gene. We identified the transcriptional start sites of each rrn operon (a G) and found that transcription of all rrn operons from their P1 promoters was drastically reduced in the relA mutant while this was almost completely restored in the relA yjbM ywaC triple mutant. Taken together with previous results showing that the intracellular GTP concentration was reduced in the relA mutant while it was restored in the triple mutant, it seems likely that continuous (p)ppGpp synthesis by YjbM and/or YwaC at a basal level causes a decrease in the amounts of intracellular GTP.

* Corresponding author. Mailing address: College of Science, Rikkyo University, Toshima-ku Nishi-ikebukuro 3-34-1, Tokyo 171-8501, Japan. Phone and fax: 81 3 3985 2401. E-mail: kawamura{at}rikkyo.ne.jp

{triangledown} Published ahead of print on 15 May 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Y.N. and K.T. contributed equally to this work.

Journal of Bacteriology, July 2009, p. 4555-4561, Vol. 191, No. 14
0021-9193/09/$08.00+0     doi:10.1128/JB.00263-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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