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Characterization and analysis of the regulatory network involved in control of lipomycin biosynthesis in Streptomyces aureofaciens Tü117

  • Applied Genetics and Molecular Biotechnology
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Abstract

Analysis of the α-lipomycin biosynthesis gene cluster of Streptomyces aureofaciens Tü117 led to the identification of five putative regulatory genes, which are congregated into a subcluster. Analysis of the lipReg1–4 and lipX1 showed that they encode components of two-component signal transduction systems (LipReg1 and LipReg2), multiple antibiotics resistance-type regulator (LipReg3), large ATP-binding regulators of the LuxR family-type regulator (LipReg4), and small ribonuclease (LipRegX1), respectively. A combination of targeted gene disruptions, complementation experiments, lipomycin production studies, and gene expression analysis via RT-PCR suggests that all regulatory lip genes are involved in α-lipomycin production. On the basis of the obtained data, we propose that LipReg2 controls the activity of LipReg1, which in its turn govern the expression of the α-lipomycin pathway-specific regulatory gene lipReg4. The ribonuclease gene lipX1 and the transporter regulator lipReg3 appear to work independently of genes lipReg1, lipReg2, and lipReg4.

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References

  • Aigle B, Pang X, Decaris B, Leblond P (2005) Involvement of AlpV, a new member of the Streptomyces antibiotic regulatory protein family, in regulation of the duplicated type II polyketide synthase alp gene cluster in Streptomyces ambofaciens. J Bacteriol 187:2491–2500

    Article  CAS  Google Scholar 

  • Alekshun M, Levy S, Mealy T, Seaton B, Head J (2001) The crystal structure of MarR, a regulator of multiple antibiotic resistance, at 2, 3 A resolution. Nat Struct Biol 8:710–714

    Article  CAS  Google Scholar 

  • Bate N, Stratigopoulos G, Cundliffe E (2002) Differential roles of two SARP-encoding regulatory gene during tylosin biosynthesis. Mol Microbiol 43:449–458

    Article  CAS  Google Scholar 

  • Bate N, Bignell DRD, Cundliffe E (2006) Regulation of tylosin biosynthesis involving ‘SARP- helper’ activity. Mol Microbiol 62:148–156

    Article  CAS  Google Scholar 

  • Bierman M, Logan R, O’Brein K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49

    Article  CAS  Google Scholar 

  • Bihlmaier C, Welle E, Hofmann C, Welzel K, Vente A, Breitling E, Muller M, Glaser S, Bechthold A (2006) Biosynthesis of the acyclic polyene antibiotic lipomycin in Streptomyces aureofaciens Tü117. Antimicrob Agents Chemother 50:2113–2121

    Article  CAS  Google Scholar 

  • Blondelet-Rouault M-H, Weiser Y, Lebrihi A, Branny P, Pernodet J (1997) Antibiotic resistance gene cassettes derived from the U` interposon for use in Escherichia coli and Streptomyces. Gene 190:315–317

    Article  CAS  Google Scholar 

  • Bolard J (1986) How do the polyene macrolide antibiotics affect the cellular membrane properties? Biochim Biophys Acta 864:257–304

    CAS  Google Scholar 

  • Boos W, Shuman H (1998) Maltose/maltodextrin system of Escherichia coli: transport, metabolism and regulation. Microbiol Mol Biol Rev 62:204–229

    CAS  Google Scholar 

  • Chen Y, Wendt-Pienkowski E, Shen B (2008) Identification and utility of FdmR1 as a Streptomyces antibiotic regulatory protein activator for fredericamycin production in Streptomyces griseus ATCC 49344 and heterologous hosts. J Bacteriol 190:5587–5596

    Article  CAS  Google Scholar 

  • Dylan C, Jensen A (1998) Investigation of the Streptomyces clavuligerus cephamycin C gene cluster and its regulation by the CcaR protein. J Bacteriol 180:4068–4079

    Google Scholar 

  • Esser V, Elefson D (1970) Experiences with the Kirby–Bauer method of antibiotic susceptibility testing. Am J Clin Pathol 54:193–198

    CAS  Google Scholar 

  • Fernandez-Moreno M, Caballero J, Hopwood D, Malpartida F (1991) The act-cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bld-tRNA gene of Streptomyces. Cell 66:769–780

    Article  CAS  Google Scholar 

  • Flett F, Mersinias V, Smith C (1997) High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting Streptomyces. FEMS Microbiol Lett 155:223–229

    Article  CAS  Google Scholar 

  • Folcher M, Gaillard H, Nguyen L, Nguyen K, Lacroix K, Bamas-Jacques N, Rinkel M, Ch T (2001) Pleiotropic function of a Streptomyces pristinaespiralis autoregulator receptor in development, antibiotic biosynthesis, and expression of a superoxide dismutase. J Biol Chem 276:44297–44306

    Article  CAS  Google Scholar 

  • Gravenbeek M, Jones G (2008) The endonuclease activity of RNase III is required for the regulation of antibiotic production by Streptomyces coelicolor. Microbiology 154:3547–3555

    Article  CAS  Google Scholar 

  • Grkovich S, Brown M, Skurray R (2002) Regulation of bacterial drug export systems. Microbiol Mol Biol Rev 66:671–701

    Article  Google Scholar 

  • Gust B, Kieser T, Chater K (2002) PCR targeting system in Streptomyces coelicolor A3(2). The John Innes Foundation, Norwich

    Google Scholar 

  • Guzmann LM, Belin D, Carson MJ, Beckwith J (1995) Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 177:4121–4130

    Google Scholar 

  • Hopwood D (2007) How do antibiotic-producing bacteria ensure their self-resistance before antibiotic biosynthesis incapacitates them? Mol Microbiol 63:937–940

    Article  CAS  Google Scholar 

  • Horinouchi S (2007) Mining and polishing of the treasure trove in the bacterial genus Streptomyce. Biosci Biotechnol Biochem 71:283–299

    Article  CAS  Google Scholar 

  • Hutchings M, Hoskisson P, Chandra G, Buttner M (2004) Sensing and responding to diverse extracellular signals? Analysis of the sensor kinase and response regulators of Streptomyces coelicolor A3(2). Microbiol 150:2795–2806

    Article  CAS  Google Scholar 

  • Kennel D (2002) Processing endoribonucleases and mRNA degradation in bacteria. J Bacteriol 184:4645–4657

    Article  Google Scholar 

  • Kieser T, Bibb M, Buttner M, Chater K, Hopwood D (2000) Practical Streptomyces genetics. The John Innes Foundation, Norwich

    Google Scholar 

  • Köseoglu O, Kocagöz T (2004) A novel T-cloning vector system. Mikrobiyol Bul 38:239–243

    Google Scholar 

  • Lee P, Umeyma T, Horinouchi S (2002) AfsS is a target of AfsR, a transcriptional factor with ATPase activity that globally controls secondary metabolism in Streptomyces coelicolor A3(2). Mol Microbiol 43:413–1430

    Google Scholar 

  • Luzhetskyy A, Fedoryshyn M, Gromyko O, Ostash B, Rebets Y, Bechthold A, Fedorenko V (2006) IncP plasmids are most effective in mediating conjugation between Escherichia coli and Streptomyces. Genetica 42:595–601

    CAS  Google Scholar 

  • McKenzie N, Nodwell J (2007) Phosphorylated AbsA2 negatively regulates antibiotic production in Streptomyces coelicolor through interactions with pathway-specific regulatory gene promoters. J Bacteriol 189:5284–5292

    Article  CAS  Google Scholar 

  • Nuria A, Mendes M, Martin J, Aparacio J (2004) Identification of PimR as a positive regulator of pimaricin biosynthesis in Streptomyces natalensis. J Bacteriol 186:2567–2575

    Article  Google Scholar 

  • Nuria A, Santos-Aberturas J, Mendes M, Guerra S, Martin J, Aparacio J (2007) PimM, a PAS domain positive regulator of pimaricin biosynthesis in Streptomyces natalensis. J Bacteriol 153:3174–3183

    Google Scholar 

  • Otten S, Ferguson J, Hutchinson R (1995) Regulation of daunorubicin production in Streptomyces peucetius by the dnrR2 locus. J Bacteriol 177:1216–1224

    CAS  Google Scholar 

  • Pang X, Aigle B, Girardet M, Mangenot S, Pernodet L, Decaris B, Leblond P (2004) Functional angucyclin like antibiotic gene cluster in the terminal inverted repeats of Streptomyces ambofaciens liner chromosome. Antimicrob Agents Chemother 48:575–588

    Article  CAS  Google Scholar 

  • Price B, Adamidis T, Kong R, Champness W (1999) A Streptomyces coelicolor antibiotic regulatory gene, absB, encodes an RNase III homolog. J Bacteriol 181:6142–6151

    CAS  Google Scholar 

  • Sambrook J, Russell D (2001) Molecular cloning, a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Saridakis V, Shahinas D, Christendat D (2008) Structural insights on the mechanism of regulation of the MarR family of proteins: high resolution crystal structure of a transcriptional repressor from Methanobacterium thermoautotrophicum. J Mol Biol 377:655–667

    Article  CAS  Google Scholar 

  • Sello J, Buttner M (2008) The gene encoding RNase III in Streptomyces coelicolor is transcribed during exponential phase and is required for antibiotic production and for proper sporulation. J Bacteriol 190:4079–4083

    Article  CAS  Google Scholar 

  • Sevcik J, Urbanikova L, Leland P, Raines R (2002) X-ray structure of two crystalline forms of a Streptomyces ribonuclease with cytotoxic activity. J Biol Chem 277:47325–47330

    Article  CAS  Google Scholar 

  • Sevcik J, Dauter Z, Wilson K (2004) Crystal structure reveals two alternative conformations in the active site of ribonuclease Sa2. Acta Crystallogr D Biol Crystallogr D60:1198–1204

    Article  CAS  Google Scholar 

  • Sheldon P, Busarow S, Hutchinson R (2002) Mapping the DNA-binding domain and target sequences of the Streptomyces peucetius daunorubicin biosynthesis regulatory protein, DnrI. Mol Microbiol 44:449–460

    Article  CAS  Google Scholar 

  • Tahlan K, Ahn S, Sing A, Bodnaruk T, Willems A, Davidson A, Nodwell J (2007) Initiation of actinorhodin export in Streptomyces coelicolor. Mol Microbiol 63:951–961

    Article  CAS  Google Scholar 

  • Wilson D, Xue Y, Reynolds K, Sherman D (2001) Characterization and analysis of the PikD regulatory factor in the picromycin biosynthetic pathway of Streptomyces venezuelae. J Bacteriol 183:3468–3475

    Article  CAS  Google Scholar 

  • Zhang Y, Muyrers JP, Rientjes J, Stewart AF (2003) Phage annealing proteins promote oligonucleotide-directed mutagenesis in Escherichia coli and mouse ES cells. BMC Mol Biol 4:1

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the BMBF (genomic plus) grant to A.B and by the DAAD grant to L.H. (PKZ A/07/99406).

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Ivan Franko National University of L’viv, Grushevskogo st.4, L’viv, 79005, Ukraine

    Liliya Horbal, Yuriy Rebets, Maria Rabyk, Bogdan Ostash & Victor Fedorenko

  2. Department of Microbiology and Department of Pharmacy and Applied Life Sciences, Niigata University, Higashijima 265-1, 956-8603, Niigata, Japan

    Tatsunosuke Nakamura

  3. Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie und Biotechnologie Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104, Freiburg, Germany

    Andriy Luzhetskyy, Elisabeth Welle & Andreas Bechthold

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  1. Liliya Horbal
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  2. Yuriy Rebets
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  3. Maria Rabyk
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  4. Andriy Luzhetskyy
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  5. Bogdan Ostash
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  8. Victor Fedorenko
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  9. Andreas Bechthold
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Correspondence to Andreas Bechthold.

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Liliya Horbal and Yuriy Rebets equally contributed to this work.

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Horbal, L., Rebets, Y., Rabyk, M. et al. Characterization and analysis of the regulatory network involved in control of lipomycin biosynthesis in Streptomyces aureofaciens Tü117. Appl Microbiol Biotechnol 85, 1069–1079 (2010). https://doi.org/10.1007/s00253-009-2108-3

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  • DOI: https://doi.org/10.1007/s00253-009-2108-3

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