Abstract
Transcription from thechiC promoter, directing expression of the chitinase gene,chiC, inStreptomyces coelicolor, was analyzed usingxylE reporter gene and high-resolution S1-nuclease mapping. The transcription from thechiC promoter was induced by chitin, and this induction was dramatically reduced in theS. coelicolor chiR-disrupted strain. This indicated a dependence ofchiC expression upon thechiR gene encoding a response regulator protein. To investigate this relationship, theS. coelicolor ChiR was over-produced usingEscherichia coli T7 RNA polymerase expression system. However, gel mobility shift-assay with such a purified ChiR showed no binding in thechiC promoter region, which indicates a lack of specific phosphorylation ofE. coli overproduced ChiR that is necessary for DNA-binding activity of response regulators.
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References
Ausubel F.M., Brent R., Kingston R.E., Moore D.O., Seidman J.S., Smithj A., Struhl K.:Current Protocols in Molecular Biology. Wiley, New York 1995.
Bradford M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72, 248–254 (1976).
Brawner M.E., Auerbach J.I., Fornwald J.A., Rosenberg M., Taylor D.P.: Characterization ofStreptomyces promoter sequence using theEscherichia coli galactokinase gene.Gene 40, 191–201 (1985).
Delic I., Robbind P., Westpheling J.: Direct repeat sequences are implicated in the regulation of twoStreptomyces chitinase promoters that are subject to carbon catabolit control.Proc. Nat. Acad. Sci. USA 89, 1885–1889 (1992).
Fujii T., Miyashita K.: Multiple domain structure in a chitinase gene (chiC) ofStreptomyces lividans.J. Gen. Microbiol. 139, 677–686 (1993).
Ghormade V.S., Lachke S.A., Deshpande M.V.: Dimorphism inBenjaminiella poitrasii: involvement of intracellular endochitinase andN-acetylglucosaminidase activities in the yeast mycelium transition.Folia Microbiol. 45, 231–238 (2000).
Gilbert M., Morosoli R., Shareck F., Kluepfel D.: Production and secretion of proteins by streptomycetes.Crit. Rev. Biotechnol. 15, 13–39 (1995).
Hakenbeck R., Stock J.B.: Analysis of two-component signal transduction systems involved in transcriptional regulation.Meth. Enzymol. 273, 281–300 (1996).
Hopwood D.A., Bibb M.J., Chater K.F., Kieser T., Bruton C.J., Kieser H.M., Lydiate D.L., Smith C.P., Ward J.M., Schrempf H.:Genetic Manipulation of Streptomyces. A Laboratory Manual. The John Innes Foundation, Norwich (UK) 1985.
Ingram C., Brawner M., Youngman P., Westpheling J.:xylE fusions as an efficient reporter gene inStreptomyces spp.: use for study ofgalP1, a catabolite-controlled promoter.J. Bacteriol. 171, 6617–6624 (1989).
Kelemen G.H., Brown G.L., Kormanec J., Potúčková L., Chater K.F., Buttner M.J.: The position of the σ factor genes,whiG andsigF, in the hierarchy controlling the development of spore chains in the aerial hyphae ofStreptomyces coelicolor A3.Mol. Microbiol. 21, 593–603 (1996).
Kopečný J., Hodrová B.: Chitinolytic enzymes produced by ovine rumen bacteria.Folia Microbiol. 45, 465–468 (2000).
Kormanec J., Ševčíková B., Homerová D.: Cloning of a two-component regulatory system probably involved in the regulation of chitinase inStreptomyces coelicolor A3(2).Folia Microbiol. 45, 397–406 (2000).
Kormanec J., Homerová D., Ševčíková B., Řežuchová B.: A method for isolation of small DNA fragments from agarose and polyacrylamide gels.Anal. Biochem.,293, 138–139 (2001).
Kormanec J.: Analyzing the developmental expression of sigma factors with S1 nuclease mapping, pp. 481–494 in C.H. Chein (Ed.):Nuclease Methods and Protocols. Methods in Molecular Biology. Humana Press, Totowa (USA) 2001.
Lima L.H.C., De Marco J.L., Ulhoa C.J., Felix C.R.: Synthesis of aTrichoderma chitinase which affects theSclerotium rolfsii andRhizoctonia solani cell walls.Folia Microbiol. 44, 45–49 (1999).
Lingappa Y., Lockwood J.L.: Chitin media for selective isolation and culture of actinomycetes.Phytopathology 52, 317–323 (1962).
Maxam A.M., Gilbert W.: Sequencing end-labeled DNA with base specific chemical cleavages.Meth. Enzymol. 65, 449–560 (1980).
Miyashita K., Fujii T., Saito A.: Induction and repression of aStreptomyces lividans chitinase gene promoter in response to various carbon sources.Biosci. Biotechnol. Biochem. 64, 39–43 (2000).
Ni X., Westpheling J.: Direct repeat sequences in theStreptomyces chitinase-63 promoter direct both glucose and chitin induction.Proc. Nat. Acad. Sci. USA 94, 13116–13121 (1997).
Robbins P.W., Oberbye K., Albright C., Benfield B., Pero J.: Cloning and high-level expression of chitinase-encoding gene ofStreptomyces plicatus.Gene 111, 69–76 (1992).
Saito A., Fujii T., Yoneyama T., Redenbach K., Ohno T., Watanabe T., Miyashita K.: High-multiplicity of chitinase genes inStreptomyces coelicolor A3(2).Biosci. Biotechnol. Biochem. 63, 710–818 (1999).
Saito A., Ishizaka M., Francisco P.B. Jr.,Fujii T., Miyashita K.: Transcriptional co-regulation of five chitinase genes scattered on theStreptomyces coelicolor A3(2) chromosome.Microbiology 146, 2937–2946 (2000).
Šimůnek J., Hodrová B., Bartoňová H., Kopečný J.: Chitinolytic bacteria of the mammal digestive tract.Folia Microbiol. 46, 76–78 (2001).
Tsujibo H., Hatano N., Endo H., Miyamoto K., Inamori Y.: Purification and characterization of a thermostable chitinase fromStreptomyces thermoviolaceus OPC-520 and cloning of the encoding gene.Biosci. Biotechnol. Biochem. 64, 96–102 (2000).
Tsujibo H., Hatano N., Okamoto T., Endo H., Miyamoto K., Inamori Y.: Synthesis of chitinase inStreptomyces thermoviolaceus is regulated by a two-component sensor-regulator system.FEMS Microbiol. Lett. 181, 83–90 (1999).
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Homerová, D., Knirschová, R. & Kormanec, J. Response regulator ChiR regulates expression of chitinase gene,chiC, inStreptomyces coelicolor . Folia Microbiol 47, 499–505 (2002). https://doi.org/10.1007/BF02818788
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DOI: https://doi.org/10.1007/BF02818788