1887

Abstract

HTA426, a thermophilic Gram-positive bacterium, feeds on inositol as its sole carbon source, and an gene cluster required for inositol catabolism has been postulated with reference to the genes in . The gene cluster of comprises two tandem operons induced in the presence of inositol; however, the mechanism underlying this induction remains unclear. is known to be involved in the regulation of encoding -inositol dehydrogenase, and its homologue in HTA426 was found two genes upstream of the first gene () of the gene cluster and was termed in . When was inactivated in , not only cellular -inositol dehydrogenase activity due to expression but also the transcription of the two operons became constitutive. IolQ was produced and purified as a C-terminal histidine (His)-tagged fusion protein in and subjected to an gel electrophoresis mobility shift assay to examine its DNA-binding property. It was observed that IolQ bound to the DNA fragments containing each of the two promoter regions and that DNA binding was antagonized by -inositol. Moreover, DNase I footprinting analyses identified two tandem binding sites of IolQ within each of the promoter regions. By comparing the sequences of the binding sites, a consensus sequence for IolQ binding was deduced to form a palindrome of 5′-RGWAAGCGCTTSCY-3′ (where R=A or G, W=A or T, S=G or C, and Y=C or T). IolQ functions as a transcriptional repressor regulating the induction of the two operons responding to -inositol.

Funding
This study was supported by the:
  • Japan Society for the Promotion of Science (Award KAKENHI (18H02128))
    • Principle Award Recipient: Ken-ichiYoshida
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2020-12-15
2021-07-29
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References

  1. Yoshida KI, Aoyama D, Ishio I, Shibayama T, Fujita Y. Organization and transcription of the myo-inositol operon, iol, of Bacillus subtilis . J Bacteriol 1997; 179:4591–4598 [View Article][PubMed]
    [Google Scholar]
  2. Yoshida KI, Shibayama T, Aoyama D, Fujita Y. Interaction of a repressor and its binding sites for regulation of the Bacillus subtilis iol divergon. J Mol Biol 1999; 285:917–929 [View Article][PubMed]
    [Google Scholar]
  3. Kang DM, Michon C, Morinaga T, Tanaka K, Takenaka S et al. Bacillus subtilis IolQ (DegA) is a transcriptional repressor of iolX encoding NAD+-dependent scyllo-inositol dehydrogenase. BMC Microbiol 2017; 17:154 [View Article][PubMed]
    [Google Scholar]
  4. Yoshida K, Yamamoto Y, Omae K, Yamamoto M, Fujita Y. Identification of two myo-inositol transporter genes of Bacillus subtilis . J Bacteriol 2002; 184:983–991 [View Article][PubMed]
    [Google Scholar]
  5. Kang DM, Tanaka K, Takenaka S, Ishikawa S, Yoshida K. Bacillus subtilis iolU encodes an additional NADP+-dependent scyllo-inositol dehydrogenase. Biosci Biotechnol Biochem 2017; 81:1026–1032 [View Article][PubMed]
    [Google Scholar]
  6. Morinaga T, Ashida H, Yoshida K. Identification of two scyllo-inositol dehydrogenases in Bacillus subtilis . Microbiology 2010; 156:1538–1546 [View Article][PubMed]
    [Google Scholar]
  7. Yoshida K, Yamaguchi M, Ikeda H, Omae K, Tsurusaki K et al. The fifth gene of the iol operon of Bacillus subtilis, iolE, encodes 2-keto-myo-inositol dehydratase. Microbiology 2004; 150:571–580 [View Article][PubMed]
    [Google Scholar]
  8. Yoshida K, Yamaguchi M, Morinaga T, Kinehara M, Ikeuchi M et al. myo-Inositol catabolism in Bacillus subtilis . J Biol Chem 2008; 283:10415–10424 [View Article][PubMed]
    [Google Scholar]
  9. Kohler PR, Choong EL, Rossbach S. The RpiR-like repressor IolR regulates inositol catabolism in Sinorhizobium meliloti . J Bacteriol 2011; 193:5155–5163 [View Article][PubMed]
    [Google Scholar]
  10. Kröger C, Fuchs TM. Characterization of the myo-inositol utilization island of Salmonella enterica serovar Typhimurium. J Bacteriol 2009; 191:545–554 [View Article][PubMed]
    [Google Scholar]
  11. Rothhardt JE, Kröger C, Broadley SP, Fuchs TM. The orphan regulator ReiD of Salmonella enterica is essential for myo-inositol utilization. Mol Microbiol 2014; 94:700–712 [View Article][PubMed]
    [Google Scholar]
  12. Takami H, Inoue A, Fuji F, Horikoshi K. Microbial flora in the deepest sea mud of the Mariana Trench. FEMS Microbiol Lett 1997; 152:279–285 [View Article][PubMed]
    [Google Scholar]
  13. Takami H, Nishi S, Lu J, Shimamura S, Takaki Y. Genomic characterization of thermophilic Geobacillus species isolated from the deepest sea mud of the Mariana Trench. Extremophiles 2004; 8:351–356 [View Article][PubMed]
    [Google Scholar]
  14. Suzuki H, Yoshida K. Genetic transformation of Geobacillus kaustophilus HTA426 by conjugative transfer of host-mimicking plasmids. J Microbiol Biotechnol 2012; 22:1279–1287 [View Article][PubMed]
    [Google Scholar]
  15. Suzuki H, Murakami A, Yoshida K. Counterselection system for Geobacillus kaustophilus HTA426 through disruption of pyrF and pyrR . Appl Environ Microbiol 2012; 78:7376–7383 [View Article][PubMed]
    [Google Scholar]
  16. Miyano M, Tanaka K, Ishikawa S, Mori K, Miguel-Arribas A et al. A novel method for transforming the thermophilic bacterium Geobacillus kaustophilus . Microb Cell Fact 2018; 17:127 [View Article][PubMed]
    [Google Scholar]
  17. Yoshida K-I, Sanbongi A, Murakami A, Suzuki H, Takenaka S et al. Three inositol dehydrogenases involved in utilization and interconversion of inositol stereoisomers in a thermophile, Geobacillus kaustophilus HTA426. Microbiology 2012; 158:1942–1952 [View Article][PubMed]
    [Google Scholar]
  18. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z et al. Gapped blast and PSI-blast: a new generation of protein database search programs. Nucleic Acids Res 1997; 25:3389–3402 [View Article][PubMed]
    [Google Scholar]
  19. Yoshida K, Yamaguchi M, Morinaga T, Ikeuchi M, Kinehara M et al. Genetic modification of Bacillus subtilis for production of D-chiro-inositol, an investigational drug candidate for treatment of type 2 diabetes and polycystic ovary syndrome. Appl Environ Microbiol 2006; 72:1310–1315 [View Article][PubMed]
    [Google Scholar]
  20. Yamaoka M, Osawa S, Morinaga T, Takenaka S, Yoshida K. A cell factory of Bacillus subtilis engineered for the simple bioconversion of myo-inositol to scyllo-inositol, a potential therapeutic agent for Alzheimer's disease. Microb Cell Fact 2011; 10:69 [View Article][PubMed]
    [Google Scholar]
  21. Herrou J, Crosson S. Myo-Inositol and D-ribose ligand discrimination in an ABC periplasmic binding protein. J Bacteriol 2013; 195:2379–2388 [View Article][PubMed]
    [Google Scholar]
  22. Yoshida K, Kobayashi K, Miwa Y, Kang CM, Matsunaga M et al. Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis . Nucleic Acids Res 2001; 29:683–692 [View Article][PubMed]
    [Google Scholar]
  23. Kawsar HI, Ohtani K, Okumura K, Hayashi H, Shimizu T. Organization and transcriptional regulation of myo-inositol operon in Clostridium perfringens. FEMS Microbiol Lett 2004; 235:289–295 [View Article][PubMed]
    [Google Scholar]
  24. Yebra MJ, Zúñiga M, Beaufils S, Pérez-Martínez G, Deutscher J et al. Identification of a gene cluster enabling Lactobacillus casei BL23 to utilize myo-inositol. Appl Environ Microbiol 2007; 73:3850–3858 [View Article][PubMed]
    [Google Scholar]
  25. Kohler PR, Zheng JY, Schoffers E, Rossbach S. Inositol catabolism, a key pathway in Sinorhizobium meliloti for competitive host nodulation. Appl Environ Microbiol 2010; 76:7972–7980 [View Article][PubMed]
    [Google Scholar]
  26. Fujita Y. Carbon catabolite control of the metabolic network in Bacillus subtilis . Biosci Biotechnol Biochem 2009; 73:45–59 [View Article][PubMed]
    [Google Scholar]
  27. Krings E, Krumbach K, Bathe B, Kelle R, Wendisch VF et al. Characterization of myo-inositol utilization by Corynebacterium glutamicum: the stimulon, identification of transporters, and influence on L-lysine formation. J Bacteriol 2006; 188:8054–8061 [View Article][PubMed]
    [Google Scholar]
  28. Lindner SN, Seibold GM, Henrich A, Kramer R, Wendisch VF. Phosphotransferase system-independent glucose utilization in Corynebacterium glutamicum by inositol permeases and glucokinases. Appl Environ Microbiol 2011; 77:3571–3581 [View Article][PubMed]
    [Google Scholar]
  29. White RH, Miller SL. Inositol isomers: occurrence in marine sediments. Science 1976; 193:885–886 [View Article][PubMed]
    [Google Scholar]
  30. Yancey PH. Compatible and counteracting solutes: protecting cells from the dead sea to the deep sea. Sci Prog 2004; 87:1–24 [View Article][PubMed]
    [Google Scholar]
  31. Yancey PH. Organic osmolytes as compatible, metabolic and counteracting cytoprotectants in high osmolarity and other stresses. J Exp Biol 2005; 208:2819–2830 [View Article][PubMed]
    [Google Scholar]
  32. Yancey PH, stress W. Osmolytes and proteins. Amer Zool 2001; 41:699–709
    [Google Scholar]
  33. Townsend M, Cleary JP, Mehta T, Hofmeister J, Lesne S et al. Orally available compound prevents deficits in memory caused by the Alzheimer amyloid-beta oligomers. Ann Neurol 2006; 60:668–676 [View Article][PubMed]
    [Google Scholar]
  34. McLaurin J, Kierstead ME, Brown ME, Hawkes CA, Lambermon MH et al. Cyclohexanehexol inhibitors of Abeta aggregation prevent and reverse Alzheimer phenotype in a mouse model. Nat Med 2006; 12:801–808 [View Article][PubMed]
    [Google Scholar]
  35. Matthews KS, Nichols JC. Lactose repressor protein: functional properties and structure. Prog Nucleic Acid Res Mol Biol 1998; 58:127–164 [View Article][PubMed]
    [Google Scholar]
  36. Oehler S, Eismann ER, Krämer H, Müller-Hill B. The three operators of the lac operon cooperate in repression. Embo J 1990; 9:973–979 [View Article][PubMed]
    [Google Scholar]
  37. Swint-Kruse L, Matthews KS. Allostery in the LacI/GalR family: variations on a theme. Curr Opin Microbiol 2009; 12:129–137 [View Article][PubMed]
    [Google Scholar]
  38. Hamoen LW, Venema G, Kuipers OP. Controlling competence in Bacillus subtilis: shared use of regulators. Microbiology 2003; 149:9–17 [View Article][PubMed]
    [Google Scholar]
  39. Suzuki H, Kobayashi J, Wada K, Furukawa M, Doi K. Thermoadaptation-directed enzyme evolution in an error-prone thermophile derived from Geobacillus kaustophilus HTA426. Appl Environ Microbiol 2015; 81:149–158 [View Article][PubMed]
    [Google Scholar]
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