Abstract
Streptomyces cyanogenus S136 is the only known producer of landomycin A (LaA), one of the largest glycosylated angucycline antibiotics possessing strong antiproliferative properties. There is rising interest in elucidation of mechanisms of action of landomycins, which, in turn, requires access to large quantities of the pure compounds. Overproduction of LaA has been achieved in the past through manipulation of cluster-situated regulatory genes. However, other components of the LaA biosynthetic regulatory network remain unknown. To fill this gap, we elucidated the contribution of AdpA family pleiotropic regulators in landomycin production via expression of adpA genes of different origins in S. cyanogenus S136. Overexpression of the native S. cyanogenus S136 adpA ortholog had no effect on landomycin titers. In the same time, expression of several heterologous adpA genes led to significantly increased landomycin production under different cultivation conditions. Hence, heterologous adpA genes are a useful tool to enhance or activate landomycin production by S. cyanogenus. Our ongoing research effort is focused on identification of mutations that render S. cyanogenus AdpA nonfunctional.
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References
Ahmed Y, Rebets Y, Tokovenko B, Brötz E, Luzhetskyy A (2017) Identification of butenolide regulatory system controlling secondary metabolism in Streptomyces albus J1074. Sci Rep 7(1):9784. https://doi.org/10.1038/s41598-017-10316-y
Bailey TL, Boden M, Buske FA, Frith M, Grant CE, Clementi L, Ren J, Li WW, Noble WS (2009) MEME SUITE: tools for motif discovery and searching. Nucleic Acids Res 37(Web Server issue):W202–W208. https://doi.org/10.1093/nar/gkp335
Claesen J, Bibb MJ (2011) Biosynthesis and regulation of grisemycin, a new member of the linaridin family of ribosomally synthesized peptides produced by Streptomyces griseus IFO 13350. J Bacteriol 193(10):2510–2516. https://doi.org/10.1128/JB.00171-11
Du YL, Li SZ, Zhou Z, Chen SF, Fan WM, Li YQ (2011) The pleiotropic regulator AdpAch is required for natamycin biosynthesis and morphological differentiation in Streptomyces chattanoogensis. Microbiology 157(Pt 5):1300–1311. https://doi.org/10.1099/mic.0.046607-0
Henkel T, Rohr J, Beale JM, Schwenen L (1990) Landomycins, new angucycline antibiotics from Streptomyces sp. I. Structural studies on landomycins A-D. J Antibiot 43(5):492–503
Higashi T, Iwasaki Y, Ohnishi Y, Horinouchi S (2007) A-factor and phosphate depletion signals are transmitted to the grixazone biosynthesis genes via the pathway-specific transcriptional activator GriR. J Bacteriol 189(9):3515–3524. https://doi.org/10.1128/JB.00055-07
Higo A, Hara H, Horinouchi S, Ohnishi Y (2012) Genome-wide distribution of AdpA, a global regulator for secondary metabolism and morphological differentiation in Streptomyces, revealed the extent and complexity of the AdpA regulatory network. DNA Res 19(3):259–273. https://doi.org/10.1093/dnares/dss010
Horbal L, Fedorenko V, Luzhetskyy A (2014) Novel and tightly regulated resorcinol and cumate-inducible expression systems for Streptomyces and other actinobacteria. Appl Microbiol Biotechnol 98(20):8641–8655. https://doi.org/10.1007/s00253-014-5918-x
Kharel MK, Pahari P, Shepherd MD, Tibrewal N, Nybo SE, Shaaban KA, Rohr J (2012) Angucyclines: biosynthesis, mode-of-action, new natural products, and synthesis. Nat Prod Rep 29(2):264–325. https://doi.org/10.1039/c1np00068c
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. John Innes Foundation, Norwich
Korynevska A, Heffeter P, Matselyukh B, Elbling L, Micksche M, Stoika R, Berger W (2007) Mechanisms underlying the anticancer activities of the angucycline landomycin E. Biochem Pharmacol 74(12):1713–1726. https://doi.org/10.1016/j.bcp.2007.08.026
Kuzniar A, van Ham RC, Pongor S, Leunissen JA (2008) The quest for orthologs: finding the corresponding gene across genomes. Trends Genet 24(11):539–551. https://doi.org/10.1016/j.tig.2008.08.009
Lehka LV, Panchuk RR, Berger W, Rohr J, Stoika RS (2015) The role of reactive oxygen species in tumor cells apoptosis induced by landomycin A. Ukr Biochem J 87(5):72–82
Liu G, Chater KF, Chandra G, Niu G, Tan H (2013) Molecular regulation of antibiotic biosynthesis in Streptomyces. Microbiol Mol Biol Rev 77(1):112–143. https://doi.org/10.1128/MMBR.00054-12
López-García MT, Santamarta I, Liras P (2010) Morphological differentiation and clavulanic acid formation are affected in a Streptomyces clavuligerus adpA-deleted mutant. Microbiology 156(8):2354–2365. https://doi.org/10.1099/mic.0.035956-0
Makitrynskyy R, Ostash B, Tsypik O, Rebets Y, Doud E, Meredith T, Luzhetskyy A, Bechthold A, Walker S, Fedorenko V (2013) Pleiotropic regulatory genes bldA, adpA and absB are implicated in production of phosphoglycolipid antibiotic moenomycin. Open Biol 3(10):130121. https://doi.org/10.1098/rsob.130121
Martín JF, Liras P (2010) Engineering of regulatory cascades and networks controlling antibiotic biosynthesis in Streptomyces. Curr Opin Microbiol 13(3):263–273. https://doi.org/10.1016/j.mib.2010.02.008
Metzker ML (2010) Sequencing technologies - the next generation. Nat Rev Genet 11(1):31–46. https://doi.org/10.1038/nrg2626
Myronovskyi M, Brötz E, Rosenkränzer B, Manderscheid N, Tokovenko B, Rebets Y, Luzhetskyy A (2016) Generation of new compounds through unbalanced transcription of landomycin A cluster. Appl Microbiol Biotechnol 100(21):9175–9186. https://doi.org/10.1007/s00253-016-7721-3
Ohnishi Y, Yamazaki H, Kato JY, Tomono A, Horinouchi S (2005) AdpA, a central transcriptional regulator in the A-factor regulatory cascade that leads to morphological development and secondary metabolism in Streptomyces griseus. Biosci Biotechnol Biochem 69(3):431–439. https://doi.org/10.1271/bbb.69.431
Ostash B, Rix U, Rix LL, Liu T, Lombo F, Luzhetskyy A, Gromyko O, Wang C, Braña AF, Méndez C, Salas JA, Fedorenko V, Rohr J (2004) Generation of new landomycins by combinatorial biosynthetic manipulation of the LndGT4 gene of the landomycin E cluster in S. globisporus. Chem Biol 11(4):547–555. https://doi.org/10.1016/j.chembiol.2004.03.011
Ostash I, Ostash B, Luzhetskyy A, Bechthold A, Walker S, Fedorenko V (2008) Coordination of export and glycosylation of landomycins in Streptomyces cyanogenus S136. FEMS Microbiol Lett 285(2):195–202. https://doi.org/10.1111/j.1574-6968.2008.01225.x
Ostash B, Korynevska A, Stoika R, Fedorenko V (2009) Chemistry and biology of landomycins, an expanding family of polyketide natural products. Mini Rev Med Chem 9(9):1040–1051
Pan Y, Liu G, Yang H, Tian Y, Tan H (2009) The pleiotropic regulator AdpA-L directly controls the pathway-specific activator of nikkomycin biosynthesis in Streptomyces ansochromogenes. Mol Microbiol 72(3):710–723. https://doi.org/10.1111/j.1365-2958.2009.06681.x
Panchuk RR, Lehka LV, Terenzi A, Matselyukh BP, Rohr J, Jha AK, Downey T, Kril IJ, Herbacek I, van Schoonhoven S, Heffeter P, Stoika RS, Berger W (2017) Rapid generation of hydrogen peroxide contributes to the complex cell death induction by the angucycline antibiotic landomycin E. Free Radic Biol Med 106:134–147. https://doi.org/10.1016/j.freeradbiomed.2017.02.024
Rabyk M, Yushchuk O, Rokytskyy I, Anisimova M, Ostash B (2018) Genomic insights into evolution of AdpA family master regulators of morphological differentiation and secondary metabolism in Streptomyces. J Mol Evol 86:204–215. https://doi.org/10.1007/s00239-018-9834-z
Rebets Y, Ostash B, Luzhetskyy A, Hoffmeister D, Brana A, Mendez C, Salas JA, Bechthold A, Fedorenko V (2003) Production of landomycins in Streptomyces globisporus 1912 and S. cyanogenus S136 is regulated by genes encoding putative transcriptional activators. FEMS Microbiol Lett 222(1):149–153
Rebets Y, Dutko L, Ostash B, Luzhetskyy A, Kulachkovskyy O, Yamaguchi T, Nakamura T, Bechthold A, Fedorenko V (2008) Function of lanI in regulation of landomycin A biosynthesis in Streptomyces cyanogenus S136 and cross-complementation studies with Streptomyces antibiotic regulatory proteins encoding genes. Arch Microbiol 189(2):111–120. https://doi.org/10.1007/s00203-007-0299-5
Sambrook J, Russell DW (2001) Molecular cloning, a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New-York
Takano E, Tao M, Long F, Bibb MJ, Wang L, Li W, Buttner MJ, Bibb MJ, Deng ZX, Chater KF (2003) A rare leucine codon in adpA is implicated in the morphological defect of bldA mutants of Streptomyces coelicolor. Mol Microbiol 50(2):475–486
Wang J, Schully KL, Pettis GS (2009) Growth-regulated expression of a bacteriocin, produced by the sweet potato pathogen Streptomyces ipomoeae, that exhibits interstrain inhibition. Appl Environ Microbiol 75(5):1236–1242. https://doi.org/10.1128/AEM.01598-08
Xu J, Zhang J, Zhuo J, Li Y, Tian Y, Tan H (2017) Activation and molecular mechanism of a cryptic oviedomycin biosynthetic gene cluster via the disruption of a global regulatory gene adpA in Streptomyces ansochromogenes. J Biol Chem 292(48):19708–19720. https://doi.org/10.1074/jbc.M117.809145
Yu P, Bu QT, Tang YL, Mao XM, Li YQ (2018) Bidirectional regulation of AdpA(ch) in controlling the expression of scnRI and scnRII in the natamycin biosynthesis of Streptomyces chattanoogensis L10. Front Microbiol 157:1300–1311. https://doi.org/10.3389/fmicb.2018.00316
Yao MD, Ohtsuka J, Nagata K, Miyazono KI, Zhi Y, Ohnishi Y, Tanokura M (2013) Complex structure of the DNA-binding domain of AdpA, the global transcription factor in Streptomyces griseus, and a target duplex DNA reveals the structural basis of its tolerant DNA sequence specificity. J Biol Chem 288:31019–31029. https://doi.org/10.1074/jbc.M113.473611
Acknowledgements
We thank Simon Shaw (Technical University of Denmark, Copenhagen) for careful proofreading of the manuscript.
Funding
This work was supported by grants Bg-46F (to V. F.) and M/26-2018 and Bg-41Nr (to B.O.) from the Ministry of Education and Science of Ukraine.
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Yushchuk, O., Ostash, I., Vlasiuk, I. et al. Heterologous AdpA transcription factors enhance landomycin production in Streptomyces cyanogenus S136 under a broad range of growth conditions. Appl Microbiol Biotechnol 102, 8419–8428 (2018). https://doi.org/10.1007/s00253-018-9249-1
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DOI: https://doi.org/10.1007/s00253-018-9249-1