Expression of a small (p)ppGpp synthetase, YwaC, in the (p)ppGpp0 mutant of Bacillus subtilis triggers YvyD-dependent dimerization of ribosome

Kazumi Tagami, Hideaki Nanamiya, Yuka Kazo, Marie Maehashi, Shota Suzuki, Eri Namba, Masahiro Hoshiya, Ryo Hanai, Yuzuru Tozawa, Takuya Morimoto, Naotake Ogasawara, Yasushi Kageyama, Katsutoshi Ara, Katsuya Ozaki, Masaki Yoshida, Haruko Kuroiwa, Tsuneyoshi Kuroiwa, Yoshiaki Ohashi, Fujio Kawamura

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

To elucidate the biological functions of small (p)ppGpp synthetases YjbM and YwaC of Bacillus subtilis, we constructed RIK1059 and RIK1066 strains carrying isopropyl-β-D-thiogalactopyranoside (IPTG) inducible yjbM and ywaC genes, respectively, in the ΔrelA ΔyjbM ΔywaC triple mutant background. While the uninduced and IPTG-induced RIK1059 cells grew similarly in LB medium, the growth of RIK1066 cells was arrested following the addition of IPTG during the early exponential growth phase. Induction of YwaC expression by IPTG also severely decreased the intracellular GTP level and drastically altered the transcriptional profile in RIK1066 cells. Sucrose density gradient centrifugation analysis of the ribosomal fractions prepared from the IPTG-induced RIK1066 cells revealed three peaks corresponding to 30S, 50S, and 70S ribosome particles, and also an extra peak. Electron microscope studies revealed that the extra peak fraction contained dimers of 70S ribosomes, which were similar to the Escherichia coli 100S ribosomes. Proteomic analysis revealed that the 70S dimer contained an extra protein, YvyD, in addition to those found in the 70S ribosome. Accordingly, strain resulting from the disruption of the yvyD gene in the RIK1066 cells was unable to form 70S dimers following IPTG induction, indicating that YvyD is required for the formation of these dimers in B. subtilis.

Original languageEnglish
Pages (from-to)115-134
Number of pages20
JournalMicrobiologyOpen
Volume1
Issue number2
DOIs
Publication statusPublished - 2012 Jun 1
Externally publishedYes

Keywords

  • (p)ppGpp
  • (p)ppGpp synthetase
  • Bacillus subtilis
  • Ribosome dimerization
  • Stringent response
  • YvyD

ASJC Scopus subject areas

  • Microbiology

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  • Cite this

    Tagami, K., Nanamiya, H., Kazo, Y., Maehashi, M., Suzuki, S., Namba, E., Hoshiya, M., Hanai, R., Tozawa, Y., Morimoto, T., Ogasawara, N., Kageyama, Y., Ara, K., Ozaki, K., Yoshida, M., Kuroiwa, H., Kuroiwa, T., Ohashi, Y., & Kawamura, F. (2012). Expression of a small (p)ppGpp synthetase, YwaC, in the (p)ppGpp0 mutant of Bacillus subtilis triggers YvyD-dependent dimerization of ribosome. MicrobiologyOpen, 1(2), 115-134. https://doi.org/10.1002/mbo3.16