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Volume 158, Issue 10

Three pathway-specific regulators control streptolydigin biosynthesis in Free

Abstract

The streptolydigin biosynthetic gene cluster from NRRL 2433 contains three putative regulatory genes, , and , encoding proteins belonging to TetR and LuxR transcriptional regulator families and ATP/GTP-binding proteins of DNA and RNA helicase superfamily I, respectively. Inactivation of or resulted in the abolition of streptolydigin production, suggesting that these proteins are pathway-specific positive regulators. In the case of the mutant, low amounts of streptolydigin C were produced instead of streptolydigin. RT-PCR transcription analysis of streptolydigin biosynthesis genes revealed a hierarchical regulation process. SlgY was found to control the expression of the regulator . SlgR2 regulates the expression of structural genes involved in the formation of the streptolydigin bicyclic ketal moiety, incorporation and processing of 3-methylaspartate, and the regulator . On the other hand, SlgR1 controls the expression of –, involved in the conversion of glutamate to 3-methylaspartate, and putative glycoside hydrolases and . Ectopic expression of , and regulatory genes in led to considerable increases in streptolydigin yields, 18-, 11- and 8.5-fold, respectively. Ectopic expression of in an mutant led to a 14-fold increase of streptolydigin C yields, while no effect was observed to result from expression of .

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2012-10-01
2024-04-19
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References

  1. Bérdy J. ( 2005). Bioactive microbial metabolites. J Antibiot (Tokyo) 58:1–26 [View Article][PubMed]
     [Google Scholar]
  2. Bibb M. J. ( 2005). Regulation of secondary metabolism in streptomycetes. Curr Opin Microbiol 8:208–215 [View Article][PubMed]
     [Google Scholar]
  3. Bihlmaier C., Welle E., Hofmann C., Welzel K., Vente A., Breitling E., Müller M., Glaser S., Bechthold A. ( 2006). Biosynthetic gene cluster for the polyenoyltetramic acid alpha-lipomycin. Antimicrob Agents Chemother 50:2113–2121 [View Article][PubMed]
     [Google Scholar]
  4. Butler A. R., Bate N., Kiehl D. E., Kirst H. A., Cundliffe E. ( 2002). Genetic engineering of aminodeoxyhexose biosynthesis in Streptomyces fradiae . Nat Biotechnol 20:713–716 [View Article][PubMed]
     [Google Scholar]
  5. Carlson J. C., Fortman J. L., Anzai Y., Li S., Burr D. A., Sherman D. H. ( 2010). Identification of the tirandamycin biosynthetic gene cluster from Streptomyces sp. 307-9. ChemBioChem 11:564–572 [View Article][PubMed]
     [Google Scholar]
  6. Chater K. F. ( 1998). Taking a genetic scalpel to the Streptomyces colony. Microbiology 144:1465–1478 [View Article]
     [Google Scholar]
  7. Chater K. F., Chandra G. ( 2008). The use of the rare UUA codon to define “expression space” for genes involved in secondary metabolism, development and environmental adaptation in Streptomyces . J Microbiol 46:1–11 [View Article][PubMed]
     [Google Scholar]
  8. Chen Y., Smanski M. J., Shen B. ( 2010). Improvement of secondary metabolite production in Streptomyces by manipulating pathway regulation. Appl Microbiol Biotechnol 86:19–25 [View Article][PubMed]
     [Google Scholar]
  9. Cundliffe E. ( 2008). Control of tylosin biosynthesis in Streptomyces fradiae . J Microbiol Biotechnol 18:1485–1491[PubMed]
     [Google Scholar]
  10. DiCioccio R. A., Srivastava B. I. ( 1976). Selective inhibition of terminal deoxynucleotidyl transferase from leukemic cells by streptolydigin. Biochem Biophys Res Commun 72:1343–1349 [View Article][PubMed]
     [Google Scholar]
  11. DiCioccio R. A., Srivastava B. I., Rinehart K. L. Jr, Lee V. J., Branfman A. R., Li L. H. ( 1980). Structure-activity relationship, selectivity and mode of inhibition of terminal deoxyribonucleotidyltransferase by streptolydigin analogs. Biochem Pharmacol 29:2001–2008 [View Article][PubMed]
     [Google Scholar]
  12. Fernández E., Weissbach U., Sánchez Reillo C., Braña A. F., Méndez C., Rohr J., Salas J. A. ( 1998). Identification of two genes from Streptomyces argillaceus encoding glycosyltransferases involved in transfer of a disaccharide during biosynthesis of the antitumor drug mithramycin. J Bacteriol 180:4929–4937[PubMed]
     [Google Scholar]
  13. Fernández-Moreno M. A., Caballero J. L., Hopwood D. A., Malpartida F. ( 1991). The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bldA tRNA gene of Streptomyces . Cell 66:769–780 [View Article][PubMed]
     [Google Scholar]
  14. Gómez C., Horna D. H., Olano C., Palomino-Schätzlein M., Pineda-Lucena A., Carbajo R. J., Braña A. F., Méndez C., Salas J. A. ( 2011). Amino acid precursor supply in the biosynthesis of the RNA polymerase inhibitor streptolydigin by Streptomyces lydicus . J Bacteriol 193:4214–4223 [View Article][PubMed]
     [Google Scholar]
  15. Gómez C., Olano C., Palomino-Schätzlein M., Pineda-Lucena A., Carbajo R. J., Braña A. F., Méndez C., Salas J. A. ( 2012a). Novel compounds produced by Streptomyces lydicus NRRL 2433 engineered mutants altered in the biosynthesis of streptolydigin. J Antibiot 65:341–348 [View Article][PubMed]
     [Google Scholar]
  16. Gómez C., Horna D. H., Olano C., Méndez C., Salas J. A. ( 2012b). Participation of putative glycoside hydrolases SlgC1 and SlgC2 in the biosynthesis of streptolydigin in Streptomyces lydicus . Microb Biotechnol [View Article][PubMed]
     [Google Scholar]
  17. He W., Lei J., Liu Y., Wang Y. ( 2008). The LuxR family members GdmRI and GdmRII are positive regulators of geldanamycin biosynthesis in Streptomyces hygroscopicus 17997. Arch Microbiol 189:501–510 [View Article][PubMed]
     [Google Scholar]
  18. Hirano S., Tanaka K., Ohnishi Y., Horinouchi S. ( 2008). Conditionally positive effect of the TetR-family transcriptional regulator AtrA on streptomycin production by Streptomyces griseus . Microbiology 154:905–914 [View Article][PubMed]
     [Google Scholar]
  19. Horbal L., Rebets Y., Rabyk M., Luzhetskyy A., Ostash B., Welle E., Nakamura T., Fedorenko V., Bechthold A. ( 2010). Characterization and analysis of the regulatory network involved in control of lipomycin biosynthesis in Streptomyces aureofaciens Tü117. Appl Microbiol Biotechnol 85:1069–1079 [View Article][PubMed]
     [Google Scholar]
  20. Horna D. H., Gómez C., Olano C., Palomino-Schätzlein M., Pineda-Lucena A., Carbajo R. J., Braña A. F., Méndez C., Salas J. A. ( 2011). Biosynthesis of the RNA polymerase inhibitor streptolydigin in Streptomyces lydicus: tailoring modification of 3-methyl-aspartate. J Bacteriol 193:2647–2651 [View Article][PubMed]
     [Google Scholar]
  21. Huang J., Shi J., Molle V., Sohlberg B., Weaver D., Bibb M. J., Karoonuthaisiri N., Lih C. J., Kao C. M. & other authors ( 2005). Cross-regulation among disparate antibiotic biosynthetic pathways of Streptomyces coelicolor . Mol Microbiol 58:1276–1287 [View Article][PubMed]
     [Google Scholar]
  22. Jiang H., Hutchinson C. R. ( 2006). Feedback regulation of doxorubicin biosynthesis in Streptomyces peucetius . Res Microbiol 157:666–674 [View Article][PubMed]
     [Google Scholar]
  23. Kieser T., Bibb M. J., Buttner M. J., Chater K. F., Hopwood D. A. ( 2000). Practical Streptomyces Genetics Norwich: The John Innes Foundation;
     [Google Scholar]
  24. Liras P., Gómez-Escribano J. P., Santamarta I. ( 2008). Regulatory mechanisms controlling antibiotic production in Streptomyces clavuligerus . J Ind Microbiol Biotechnol 35:667–676 [View Article][PubMed]
     [Google Scholar]
  25. Menéndez N., Nur-e-Alam M., Fischer C., Braña A. F., Salas J. A., Rohr J., Méndez C. ( 2006). Deoxysugar transfer during chromomycin A3 biosynthesis in Streptomyces griseus subsp. griseus: new derivatives with antitumor activity. Appl Environ Microbiol 72:167–177 [View Article][PubMed]
     [Google Scholar]
  26. Mo X., Wang Z., Wang B., Ma J., Huang H., Tian X., Zhang S., Zhang C., Ju J. ( 2011). Cloning and characterization of the biosynthetic gene cluster of the bacterial RNA polymerase inhibitor tirandamycin from marine-derived Streptomyces sp. SCSIO1666. Biochem Biophys Res Commun 406:341–347 [View Article][PubMed]
     [Google Scholar]
  27. Olano C., Wilkinson B., Sánchez C., Moss S. J., Sheridan R., Math V., Weston A. J., Braña A. F., Martin C. J., Oliynyk M. ( 2004). Biosynthesis of the angiogenesis inhibitor borrelidin by Streptomyces parvulus Tü4055: cluster analysis and assignment of functions. Chem Biol 11:87–97 [View Article][PubMed]
     [Google Scholar]
  28. Olano C., Lombó F., Méndez C., Salas J. A. ( 2008). Improving production of bioactive secondary metabolites in actinomycetes by metabolic engineering. Metab Eng 10:281–292 [View Article][PubMed]
     [Google Scholar]
  29. Olano C., Gómez C., Pérez M., Palomino M., Pineda-Lucena A., Carbajo R. J., Braña A. F., Méndez C., Salas J. A. ( 2009). Deciphering biosynthesis of the RNA polymerase inhibitor streptolydigin and generation of glycosylated derivatives. Chem Biol 16:1031–1044 [View Article][PubMed]
     [Google Scholar]
  30. Otten S. L., Olano C., Hutchinson C. R. ( 2000). The dnrO gene encodes a DNA-binding protein that regulates daunorubicin production in Streptomyces peucetius by controlling expression of the dnrN pseudo response regulator gene. Microbiology 146:1457–1468[PubMed]
     [Google Scholar]
  31. Ramos J. L., Martínez-Bueno M., Molina-Henares A. J., Terán W., Watanabe K., Zhang X., Gallegos M. T., Brennan R., Tobes R. ( 2005). The TetR family of transcriptional repressors. Microbiol Mol Biol Rev 69:326–356 [View Article][PubMed]
     [Google Scholar]
  32. Rodríguez M., Núñez L. E., Braña A. F., Méndez C., Salas J. A., Blanco G. ( 2008). Identification of transcriptional activators for thienamycin and cephamycin C biosynthetic genes within the thienamycin gene cluster from Streptomyces cattleya . Mol Microbiol 69:633–645 [View Article][PubMed]
     [Google Scholar]
  33. Sambrook J., Fritsch E. F., Maniatis T. ( 1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  34. Sánchez-Hidalgo M., Núñez L. E., Méndez C., Salas J. A. ( 2010). Involvement of the beta subunit of RNA polymerase in resistance to streptolydigin and streptovaricin in the producer organisms Streptomyces lydicus and Streptomyces spectabilis . Antimicrob Agents Chemother 54:1684–1692 [View Article][PubMed]
     [Google Scholar]
  35. Schobert R., Schlenk A. ( 2008). Tetramic and tetronic acids: an update on new derivatives and biological aspects. Bioorg Med Chem 16:4203–4221 [View Article][PubMed]
     [Google Scholar]
  36. Siebenberg S., Bapat P. M., Lantz A. E., Gust B., Heide L. ( 2010). Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates. J Biosci Bioeng 109:230–234 [View Article][PubMed]
     [Google Scholar]
  37. Temiakov D., Zenkin N., Vassylyeva M. N., Perederina A., Tahirov T. H., Kashkina E., Savkina M., Zorov S., Nikiforov V. & other authors ( 2005). Structural basis of transcription inhibition by antibiotic streptolydigin. Mol Cell 19:655–666 [View Article][PubMed]
     [Google Scholar]
  38. Tuske S., Sarafianos S. G., Wang X., Hudson B., Sineva E., Mukhopadhyay J., Birktoft J. J., Leroy O., Ismail S. & other authors ( 2005). Inhibition of bacterial RNA polymerase by streptolydigin: stabilization of a straight-bridge-helix active-center conformation. Cell 122:541–552 [View Article][PubMed]
     [Google Scholar]
  39. Tuteja N., Tuteja R. ( 2004). Prokaryotic and eukaryotic DNA helicases. Essential molecular motor proteins for cellular machinery. Eur J Biochem 271:1835–1848 [View Article][PubMed]
     [Google Scholar]
  40. Ventura M., Canchaya C., Tauch A., Chandra G., Fitzgerald G. F., Chater K. F., van Sinderen D. ( 2007). Genomics of Actinobacteria: tracing the evolutionary history of an ancient phylum. Microbiol Mol Biol Rev 71:495–548 [View Article][PubMed]
     [Google Scholar]
  41. Walker J. E., Saraste M., Runswick M. J., Gay N. J. ( 1982). Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951[PubMed]
     [Google Scholar]
  42. Wilson D. J., Xue Y., Reynolds K. A., Sherman D. H. ( 2001). Characterization and analysis of the PikD regulatory factor in the pikromycin biosynthetic pathway of Streptomyces venezuelae . J Bacteriol 183:3468–3475 [View Article][PubMed]
     [Google Scholar]
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Three pathway-specific regulators control streptolydigin biosynthesis in Streptomyces lydicus
Microbiology 158, 2504 (2012); https://doi.org/10.1099/mic.0.061325-0
/content/journal/micro/10.1099/mic.0.061325-0
/content/journal/micro/10.1099/mic.0.061325-0
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