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afsQ1-Q2-sigQ is a pleiotropic but conditionally required signal transduction system for both secondary metabolism and morphological development in Streptomyces coelicolor

  • Dan Shu
  • Lei Chen
  • Weihua Wang
  • Zhenyu Yu
  • Cong Ren
  • Weiwen Zhang
  • Sheng Yang
  • Yinhua LuEmail author
  • Weihong JiangEmail author
Applied Microbial and Cell Physiology

Abstract

Two-component system AfsQ1-Q2 of Streptomyces coelicolor was identified previously for its ability to stimulate actinorhodin (ACT) and undecylprodigiosin (RED) production in Streptomyces lividans. However, disruption of either afsQ1 or afsQ2 in S. coelicolor led to no detectable changes in secondary metabolite formation or morphogenesis. In this study, we reported that, when cultivated on defined minimal medium (MM) with glutamate as the sole nitrogen source, the afsQ mutant exhibited significantly decreased ACT, RED, and calcium-dependent antibiotic (CDA) production and rapid growth of aerial mycelium. In addition, we also found that deletion of sigQ, which is located upstream of afsQ1-Q2 and encodes a putative sigma factor, led to the precocious hyperproduction of these antibiotics and delayed formation of sporulating aerial mycelium in the same glutamate-based defined MM. Reverse-transcription polymerase chain reaction and egfp fusion analyses showed that the expression of sigQ was under control by afsQ. In addition, deletion of both afsQ-sigQ resulted in the phenotype identical to that of afsQ mutant. The results suggested that afsQ1-Q2 and sigQ worked together in the regulation of both antibiotic biosynthesis and morphological development, and sigQ might be responsible for antagonizing the function of AfsQ1-Q2 in S. coelicolor, however, in a medium-dependent manner. Moreover, the study showed that the medium-dependent regulation of antibiotic biosynthesis by AfsQ1-Q2-SigQ was through pathway-specific activator genes actII-ORF4, redD, and cdaR. The study provides new insights on regulation of antibiotic biosynthesis and morphological development in S. coelicolor.

Keywords

Morphological differentiation Sigma factor Secondary metabolism Streptomyces coelicolor Two-component system 

Notes

Acknowledgement

This work was supported by the National Natural Science Foundation of China (30770023, 30700022), the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-G-007), and the National Basic Research Program of China (2007CB707803).

Supplementary material

253_2008_1738_MOESM1_ESM.doc (40 kb)
Supplementary Table 1 Primers used in the construction of deletion mutants and gene overexpression in this study (DOC 40 kb)
253_2008_1738_MOESM2_ESM.doc (43 kb)
Supplementary Table 2 List of primers used in RT-PCR analysis (DOC 44 kb)

References

  1. Aceti DJ, Champness WC (1998) Transcriptional regulation of Streptomyces coelicolor pathway-specific antibiotic regulators by the absA and absB loci. J Bacteriol 180:3100–3106Google Scholar
  2. Adamidis T, Riggle P, Champness W (1990) Mutations in a new Streptomyces coelicolor locus which globally block antibiotic biosynthesis but not sporulation. J Bacteriol 172:2962–2969Google Scholar
  3. Anderson T, Brian P, Riggle P, Kong RQ, Champness W (1999) Genetic suppression analysis of non-antibiotic-producing mutants of the Streptomyces coelicolor absA locus. Microbiology 145:2343–2353Google Scholar
  4. Anderson TB, Brian P, Champness WC (2001) Genetic and transcriptional analysis of absA, an antibiotic gene cluster-linked two-component system that regulates multiple antibiotics in Streptomyces coelicolor. Mol Microbiol 39:553–566CrossRefGoogle Scholar
  5. Arias P, Fernandez-Moreno MA, Malpartida F (1999) Characterization of the pathway-specific positive transcriptional regulator for actinorhodin biosynthesis in Streptomyces coelicolor A3(2) as a DNA-binding protein. J Bacteriol 181:6958–6968Google Scholar
  6. Bentley SD, Chater KF, Cerdeno-Tarraga AM, Challis GL, Thomson NR, James KD, Harris DE, Quail MA, Kieser H, Harper D, Bateman A, Brown S, Chandra G, Chen CW, Collins M, Cronin A, Fraser A, Goble A, Hidalgo J, Hornsby T, Howarth S, Huang CH, Kieser T, Larke L, Murphy L, Oliver K, O’Neil S, Rabbinowitsch E, Rajandream MA, Rutherford K, Rutter S, Seeger K, Saunders D, Sharp S, Squares R, Squares S, Taylor K, Warren T, Wietzorrek A, Woodward J, Barrell BG, Parkhill J, Hopwood DA (2002) Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417:141–147CrossRefGoogle Scholar
  7. Brian P, Riggle FJ, Santos RA, Champness WC (1996) Global negative regulation of Streptomyces coelicolor antibiotic synthesis mediated by an absA-encoded putative signal transduction system. J Bacteriol 178:3221–3231Google Scholar
  8. Chakraburtty R, White J, Takano E, Bibb M (1996) Cloning, characterization and disruption of a (p)ppGpp synthetase gene (relA) of Streptomyces coelicolor A3(2). Mol Microbiol 19:357–368CrossRefGoogle Scholar
  9. Chang HM, Chen MY, Shieh YT, Bibb MJ, Chen CW (1996) The cutRS signal transduction system of Streptomyces lividans represses the biosynthesis of the polyketide antibiotic actinorhodin. Mol Microbiol 21:1075–1085Google Scholar
  10. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645CrossRefGoogle Scholar
  11. Fisher SH (1999) Regulation of nitrogen metabolism in Bacillus subtilis: vive la difference!. Mol Microbiol 32:223–232CrossRefGoogle Scholar
  12. Fisher SH, Sonenshein AL (1991) Control of carbon and nitrogen-metabolism in Bacillus subtilis. Annu Rev Microbiol 45:107–135CrossRefGoogle Scholar
  13. Gust B, Challis GL, Fowler K, Kieser T, Chater KF (2003) PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci U S A 100:1541–1546CrossRefGoogle Scholar
  14. Hoch JA (2000) Two-component and phosphorelay signal transduction. Curr Opin Microbiol 3:165–170CrossRefGoogle Scholar
  15. Hong HJ, Paget MSB, Buttner MJ (2002) A signal transduction system in Streptomyces coelicolor that activates the expression of a putative cell wall glycan operon in response to vancomycin and other cell wall-specific antibiotics. Mol Microbiol 44:1199–1211CrossRefGoogle Scholar
  16. Horinouchi S, Beppu T (1992) Autoregulatory factors and communication in Actinomycetes. Annu Rev Microbiol 46:377–398CrossRefGoogle Scholar
  17. Hutchings MI, Hoskisson PA, Chandra G, Buttner MJ (2004) Sensing and responding to diverse extracellular signals? Analysis of the sensor kinases and response regulators of Streptomyces coelicolor A3(2). Microbiology 150:2795–2806CrossRefGoogle Scholar
  18. Hutchings MI, Hong HJ, Buttner MJ (2006a) The vancomycin resistance VanRS two-component signal transduction system of Streptomyces coelicolor. Mol Microbiol 59:923–935CrossRefGoogle Scholar
  19. Hutchings MI, Hong HJ, Leibovitz E, Sutcliffe IC, Buttner MJ (2006b) The sigma(E) cell envelope stress response of Streptomyces coelicolor is influenced by a novel lipoprotein, CseA. J Bacteriol 188:7222–7229CrossRefGoogle Scholar
  20. Ikeda H, Ishikawa J, Hanamoto A, Shinose M, Kikuchi H, Shiba T, Sakaki Y, Hattori M, Omura S (2003) Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat Biotechnol 21:526–531CrossRefGoogle Scholar
  21. Ishizuka H, Horinouchi S, Kieser HM, Hopwood DA, Beppu T (1992) A putative 2-component regulatory system involved in secondary metabolism in Streptomyces spp. J Bacteriol 174:7585–7594Google Scholar
  22. Kieser T, Bibb MJ, Buttler MJ, Chater K, Hopwood D (2000) Practical Streptomyces genetics. John Innes Foundation, NorwichGoogle Scholar
  23. Lu YH, Wang WH, Shu D, Zhang WW, Chen L, Qin ZJ, Yang S, Jiang WH (2007) Characterization of a novel two-component regulatory system involved in the regulation of both actinorhodin and a type I polyketide in Streptomyces coelicolor. Appl Microbiol Biotechnol 77:625–635CrossRefGoogle Scholar
  24. Ma HT, Kendall K (1994) Cloning and analysis of a gene-cluster from Streptomyces coelicolor that causes accelerated aerial mycelium formation in Streptomyces lividans. J Bacteriol 176:3800–3811Google Scholar
  25. Maheswaran M, Forchhammer K (2003) Carbon-source-dependent nitrogen regulation in Escherichia coli is mediated through glutamine-dependent GlnB signalling. Microbiology 149:2163–2172CrossRefGoogle Scholar
  26. McKenzie NL, Nodwell JR (2007) Phosphorylated AbsA2 negatively regulates antibiotic production in Streptomyces coelicolor through interactions with pathway-specific regulatory gene promoters. J Bacteriol 189:5284–5292CrossRefGoogle Scholar
  27. Mizuno T (1997) Compilation of all genes encoding two-component phosphotransfer signal transducers in the genome of Escherichia coli. DNA Res 4:161–168CrossRefGoogle Scholar
  28. Mizuno T (2005) Two-component phosphorelay signal transduction systems in plants: from hormone responses to circadian rhythms. Biosci Biotechnol Biochem 69:2263–2276CrossRefGoogle Scholar
  29. Ohnishi Y, Ishikawa J, Hara H, Suzuki H, Ikenoya M, Ikeda H, Yamashita A, Hattori M, Horinouchi S (2008) Genome sequence of the streptomycin-producing microorganism Streptomyces griseus IFO 13350. J Bacteriol 190:4050–4060CrossRefGoogle Scholar
  30. Paget MSB, Chamberlin L, Atrih A, Foster SJ, Buttner MJ (1999a) Evidence that the extracytoplasmic function sigma factor sigma(E) is required for normal cell wall structure in Streptomyces coelicolor A3(2). J Bacteriol 181:204–211Google Scholar
  31. Paget MSB, Leibovitz E, Buttner MJ (1999b) A putative two-component signal transduction system regulates sigma(E), a sigma factor required for normal cell wall integrity in Streptomyces coelicolor A3(2). Mol Microbiol 33:97–107CrossRefGoogle Scholar
  32. Parkinson JS, Kofoid EC (1992) Communication Modules in bacterial signaling proteins. Annu Rev Genet 26:71–112CrossRefGoogle Scholar
  33. Redenbach M, Kieser HM, Denapaite D, Eichner A, Cullum J, Kinashi H, Hopwood DA (1996) A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol 21:77–96CrossRefGoogle Scholar
  34. Rodriguez-Garcia A, Barreiro C, Santos-Beneit F, Sola-Landa A, Martin JF (2007) Genome-wide transcriptomic and proteomic analysis of the primary response to phosphate limitation in Streptomyces coelicolor M145 and in a Delta phoP mutant. Proteomics 7:2410–2429CrossRefGoogle Scholar
  35. Ryding NJ, Anderson TB, Champness WC (2002) Regulation of the Streptomyces coelicolor calcium-dependent antibiotic by absA, encoding a cluster-linked two-component system. J Bacteriol 184:794–805CrossRefGoogle Scholar
  36. Sheeler NL, MacMillan SV, Nodwell JR (2005) Biochemical activities of the absA two-component system of Streptomyces coelicolor. J Bacteriol 187:687–696CrossRefGoogle Scholar
  37. Sola-Landa A, Moura RS, Martin JF (2003) The two-component PhoR-PhoP system controls both primary metabolism and secondary metabolite biosynthesis in Streptomyces lividans. Proc Natl Acad Sci U S A 100:6133–6138CrossRefGoogle Scholar
  38. Sola-Landa A, Rodriguez-Garcia A, Franco-Dominguez E, Martin JF (2005) Binding of PhoP to promoters of phosphate-regulated genes in Streptomyces coelicolor: identification of PHO boxes. Mol Microbiol 56:1373–1385CrossRefGoogle Scholar
  39. Sola-Landa A, Rodriguez-Garci A, Apel AK, Martin JF (2008) Target genes and structure of the direct repeats in the DNA-binding sequences of the response regulator PhoP in Streptomyces coelicolor. Nucleic Acids Res 36:1358–1368CrossRefGoogle Scholar
  40. Sun JH, Kelemen GH, Fernandez-Abalos JM, Bibb MJ (1999) Green fluorescent protein as a reporter for spatial and temporal gene expression in Streptomyces coelicolor A3(2). Microbiology 145:2221–2227Google Scholar
  41. Takano E, Gramajo HC, Strauch E, Andres N, White J, Bibb MJ (1992) Transcriptional regulation of the RedD transcriptional activator gene accounts for growth-phase-dependent production of the antibiotic undecylprodigiosin in Streptomyces coelicolor A3(2). Mol Microbiol 6:2797–2804CrossRefGoogle Scholar
  42. Ulrich LE, Koonin EV, Zhulin IB (2005) One-component systems dominate signal transduction in prokaryotes. Trends Microbiol 13:52–56CrossRefGoogle Scholar
  43. Umeyama T, Horinouchi S (2001) Autophosphorylation of a bacterial serine/threonine kinase, AfsK, is inhibited by KbpA, an AfsK-binding protein. J Bacteriol 183:5506–5512CrossRefGoogle Scholar
  44. Wang WW, Gao J, Chiao JS, Zhao GP, Jiang WH (2004) A novel two-component system amrB-amkB involved in the regulation of central carbohydrate metabolism in rifamycin SV-producing Amycolatopsis mediterranei U32. Curr Microbiol 48:14–19CrossRefGoogle Scholar
  45. Wei G, Jiang WH, Yang YL, Chiao JS (2004) Improvement of transformation and electroduction in avermectin high-producer, Streptomyces avermitilis. Folia Microbiol 49:399–405CrossRefGoogle Scholar
  46. White J, Bibb M (1997) bldA dependence of undecylprodigiosin production in Streptomyces coelicolor A3(2) involves a pathway-specific regulatory cascade. J Bacteriol 179:627–633Google Scholar
  47. Zhang WW, Shi L (2005) Distribution and evolution of multiple-step phosphorelay in prokaryotes: lateral domain recruitment involved in the formation of hybrid-type histidine kinases. Microbiology 151:2159–2173CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Dan Shu
    • 1
    • 2
  • Lei Chen
    • 1
    • 2
  • Weihua Wang
    • 1
    • 2
  • Zhenyu Yu
    • 1
    • 2
  • Cong Ren
    • 1
    • 2
  • Weiwen Zhang
    • 3
  • Sheng Yang
    • 1
  • Yinhua Lu
    • 1
    Email author
  • Weihong Jiang
    • 1
    Email author
  1. 1.Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiPeople’s Republic of China
  2. 2.Graduate University of the Chinese Academy of SciencesBeijingPeople’s Republic of China
  3. 3.Center for Ecogenomics, Biodesign InstituteArizona State UniversityTempeUSA

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