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. 1994 Jun;176(12):3800–3811. doi: 10.1128/jb.176.12.3800-3811.1994

Cloning and analysis of a gene cluster from Streptomyces coelicolor that causes accelerated aerial mycelium formation in Streptomyces lividans.

H Ma 1, K Kendall 1
PMCID: PMC205570  PMID: 8206859

Abstract

We describe the cloning and analysis of two overlapping DNA fragments from Streptomyces coelicolor that cause aerial mycelium to appear more rapidly than usual when introduced into Streptomyces lividans on a low-copy-number plasmid vector. Colonies of S. lividans TK64 harboring either clone produce visible aerial mycelia after only 48 h of growth, rather than the usual 72 to 96 h. From deletion and sequence analysis, this rapid aerial mycelium (Ram) phenotype appears to be due to a cluster of three genes that we have designated ramA, ramB, and ramR. Both ramA and ramB potentially encode 65-kDa proteins with homology to ATP-dependent membrane-translocating proteins. A chromosomal ramB disruption mutant of S. lividans was found to be severely defective in aerial mycelium formation. ramR could encode a 21-kDa protein with significant homology to the UhpA subset of bacterial two-component response regulator proteins. The overall organization and potential proteins encoded by the cloned DNA suggest that this is the S. coelicolor homolog of the amf gene cluster that has been shown to be important for aerial mycelium formation in Streptomyces griseus. However, despite the fact that the two regions probably have identical functions, there is relatively poor homology between the two gene clusters at the DNA sequence level.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adamidis T., Champness W. Genetic analysis of absB, a Streptomyces coelicolor locus involved in global antibiotic regulation. J Bacteriol. 1992 Jul;174(14):4622–4628. doi: 10.1128/jb.174.14.4622-4628.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adamidis T., Riggle P., Champness W. Mutations in a new Streptomyces coelicolor locus which globally block antibiotic biosynthesis but not sporulation. J Bacteriol. 1990 Jun;172(6):2962–2969. doi: 10.1128/jb.172.6.2962-2969.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baxter-Gabbard K. L. A simple method for the large-scale preparation of sucrose gradients. FEBS Lett. 1972 Jan 15;20(1):117–119. doi: 10.1016/0014-5793(72)80031-0. [DOI] [PubMed] [Google Scholar]
  4. Bibb M. J., Cohen S. N. Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet. 1982;187(2):265–277. doi: 10.1007/BF00331128. [DOI] [PubMed] [Google Scholar]
  5. Bibb M. J., Findlay P. R., Johnson M. W. The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene. 1984 Oct;30(1-3):157–166. doi: 10.1016/0378-1119(84)90116-1. [DOI] [PubMed] [Google Scholar]
  6. Blight M. A., Holland I. B. Structure and function of haemolysin B,P-glycoprotein and other members of a novel family of membrane translocators. Mol Microbiol. 1990 Jun;4(6):873–880. doi: 10.1111/j.1365-2958.1990.tb00660.x. [DOI] [PubMed] [Google Scholar]
  7. Burbulys D., Trach K. A., Hoch J. A. Initiation of sporulation in B. subtilis is controlled by a multicomponent phosphorelay. Cell. 1991 Feb 8;64(3):545–552. doi: 10.1016/0092-8674(91)90238-t. [DOI] [PubMed] [Google Scholar]
  8. Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cangelosi G. A., Martinetti G., Leigh J. A., Lee C. C., Thienes C., Theines C., Nester E. W. Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan. J Bacteriol. 1989 Mar;171(3):1609–1615. doi: 10.1128/jb.171.3.1609-1615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Champness W. C. New loci required for Streptomyces coelicolor morphological and physiological differentiation. J Bacteriol. 1988 Mar;170(3):1168–1174. doi: 10.1128/jb.170.3.1168-1174.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chater K. F. A morphological and genetic mapping study of white colony mutants of Streptomyces coelicolor. J Gen Microbiol. 1972 Aug;72(1):9–28. doi: 10.1099/00221287-72-1-9. [DOI] [PubMed] [Google Scholar]
  12. Chen C. J., Chin J. E., Ueda K., Clark D. P., Pastan I., Gottesman M. M., Roninson I. B. Internal duplication and homology with bacterial transport proteins in the mdr1 (P-glycoprotein) gene from multidrug-resistant human cells. Cell. 1986 Nov 7;47(3):381–389. doi: 10.1016/0092-8674(86)90595-7. [DOI] [PubMed] [Google Scholar]
  13. Dente L., Cesareni G., Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. doi: 10.1093/nar/11.6.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Fernández-Moreno M. A., Caballero J. L., Hopwood D. A., Malpartida F. The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bldA tRNA gene of Streptomyces. Cell. 1991 Aug 23;66(4):769–780. doi: 10.1016/0092-8674(91)90120-n. [DOI] [PubMed] [Google Scholar]
  16. Fernández-Moreno M. A., Martín-Triana A. J., Martínez E., Niemi J., Kieser H. M., Hopwood D. A., Malpartida F. abaA, a new pleiotropic regulatory locus for antibiotic production in Streptomyces coelicolor. J Bacteriol. 1992 May;174(9):2958–2967. doi: 10.1128/jb.174.9.2958-2967.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ferrari F. A., Trach K., LeCoq D., Spence J., Ferrari E., Hoch J. A. Characterization of the spo0A locus and its deduced product. Proc Natl Acad Sci U S A. 1985 May;82(9):2647–2651. doi: 10.1073/pnas.82.9.2647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Friedrich M. J., Kadner R. J. Nucleotide sequence of the uhp region of Escherichia coli. J Bacteriol. 1987 Aug;169(8):3556–3563. doi: 10.1128/jb.169.8.3556-3563.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harasym M., Zhang L. H., Chater K., Piret J. The Streptomyces coelicolor A3(2) bldB region contains at least two genes involved in morphological development. J Gen Microbiol. 1990 Aug;136(8):1543–1550. doi: 10.1099/00221287-136-8-1543. [DOI] [PubMed] [Google Scholar]
  20. Hopwood D. A., Chater K. F., Dowding J. E., Vivian A. Advances in Streptomyces coelicolor genetics. Bacteriol Rev. 1973 Sep;37(3):371–405. doi: 10.1128/br.37.3.371-405.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hopwood D. A., Kieser T., Wright H. M., Bibb M. J. Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol. 1983 Jul;129(7):2257–2269. doi: 10.1099/00221287-129-7-2257. [DOI] [PubMed] [Google Scholar]
  22. Horinouchi S., Beppu T. Autoregulatory factors and communication in actinomycetes. Annu Rev Microbiol. 1992;46:377–398. doi: 10.1146/annurev.mi.46.100192.002113. [DOI] [PubMed] [Google Scholar]
  23. Horinouchi S., Hara O., Beppu T. Cloning of a pleiotropic gene that positively controls biosynthesis of A-factor, actinorhodin, and prodigiosin in Streptomyces coelicolor A3(2) and Streptomyces lividans. J Bacteriol. 1983 Sep;155(3):1238–1248. doi: 10.1128/jb.155.3.1238-1248.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ishizuka H., Horinouchi S., Kieser H. M., Hopwood D. A., Beppu T. A putative two-component regulatory system involved in secondary metabolism in Streptomyces spp. J Bacteriol. 1992 Dec;174(23):7585–7594. doi: 10.1128/jb.174.23.7585-7594.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Karow M., Georgopoulos C. The essential Escherichia coli msbA gene, a multicopy suppressor of null mutations in the htrB gene, is related to the universally conserved family of ATP-dependent translocators. Mol Microbiol. 1993 Jan;7(1):69–79. doi: 10.1111/j.1365-2958.1993.tb01098.x. [DOI] [PubMed] [Google Scholar]
  26. Kendall K. J., Cohen S. N. Complete nucleotide sequence of the Streptomyces lividans plasmid pIJ101 and correlation of the sequence with genetic properties. J Bacteriol. 1988 Oct;170(10):4634–4651. doi: 10.1128/jb.170.10.4634-4651.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  28. Lawlor E. J., Baylis H. A., Chater K. F. Pleiotropic morphological and antibiotic deficiencies result from mutations in a gene encoding a tRNA-like product in Streptomyces coelicolor A3(2). Genes Dev. 1987 Dec;1(10):1305–1310. doi: 10.1101/gad.1.10.1305. [DOI] [PubMed] [Google Scholar]
  29. Lydiate D. J., Malpartida F., Hopwood D. A. The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors. Gene. 1985;35(3):223–235. doi: 10.1016/0378-1119(85)90001-0. [DOI] [PubMed] [Google Scholar]
  30. Merrick M. J. A morphological and genetic mapping study of bald colony mutants of Streptomyces coelicolor. J Gen Microbiol. 1976 Oct;96(2):299–315. doi: 10.1099/00221287-96-2-299. [DOI] [PubMed] [Google Scholar]
  31. Narva K. E., Feitelson J. S. Nucleotide sequence and transcriptional analysis of the redD locus of Streptomyces coelicolor A3(2). J Bacteriol. 1990 Jan;172(1):326–333. doi: 10.1128/jb.172.1.326-333.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ramakrishnan G., Newton A. FlbD of Caulobacter crescentus is a homologue of the NtrC (NRI) protein and activates sigma 54-dependent flagellar gene promoters. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2369–2373. doi: 10.1073/pnas.87.6.2369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Riordan J. R., Rommens J. M., Kerem B., Alon N., Rozmahel R., Grzelczak Z., Zielenski J., Lok S., Plavsic N., Chou J. L. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 1989 Sep 8;245(4922):1066–1073. doi: 10.1126/science.2475911. [DOI] [PubMed] [Google Scholar]
  34. Rudner D. Z., LeDeaux J. R., Ireton K., Grossman A. D. The spo0K locus of Bacillus subtilis is homologous to the oligopeptide permease locus and is required for sporulation and competence. J Bacteriol. 1991 Feb;173(4):1388–1398. doi: 10.1128/jb.173.4.1388-1398.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sanders D. A., Gillece-Castro B. L., Stock A. M., Burlingame A. L., Koshland D. E., Jr Identification of the site of phosphorylation of the chemotaxis response regulator protein, CheY. J Biol Chem. 1989 Dec 25;264(36):21770–21778. [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schrempf H. Deletion and amplification of DNA sequences in melanin-negative variants of Streptomyces reticuli. Mol Gen Genet. 1983;189(3):501–505. doi: 10.1007/BF00325917. [DOI] [PubMed] [Google Scholar]
  38. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  39. Stanfield S. W., Ielpi L., O'Brochta D., Helinski D. R., Ditta G. S. The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB. J Bacteriol. 1988 Aug;170(8):3523–3530. doi: 10.1128/jb.170.8.3523-3530.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stein D., Cohen S. N. A cloned regulatory gene of Streptomyces lividans can suppress the pigment deficiency phenotype of different developmental mutants. J Bacteriol. 1989 Apr;171(4):2258–2261. doi: 10.1128/jb.171.4.2258-2261.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stock A. M., Mottonen J. M., Stock J. B., Schutt C. E. Three-dimensional structure of CheY, the response regulator of bacterial chemotaxis. Nature. 1989 Feb 23;337(6209):745–749. doi: 10.1038/337745a0. [DOI] [PubMed] [Google Scholar]
  42. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Thompson C. J., Kieser T., Ward J. M., Hopwood D. A. Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene. 1982 Nov;20(1):51–62. doi: 10.1016/0378-1119(82)90086-5. [DOI] [PubMed] [Google Scholar]
  44. Ueda K., Miyake K., Horinouchi S., Beppu T. A gene cluster involved in aerial mycelium formation in Streptomyces griseus encodes proteins similar to the response regulators of two-component regulatory systems and membrane translocators. J Bacteriol. 1993 Apr;175(7):2006–2016. doi: 10.1128/jb.175.7.2006-2016.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Walker J. E., Saraste M., Runswick M. J., Gay N. J. 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. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wang R. C., Seror S. J., Blight M., Pratt J. M., Broome-Smith J. K., Holland I. B. Analysis of the membrane organization of an Escherichia coli protein translocator, HlyB, a member of a large family of prokaryote and eukaryote surface transport proteins. J Mol Biol. 1991 Feb 5;217(3):441–454. doi: 10.1016/0022-2836(91)90748-u. [DOI] [PubMed] [Google Scholar]
  47. Weston L. A., Kadner R. J. Role of uhp genes in expression of the Escherichia coli sugar-phosphate transport system. J Bacteriol. 1988 Aug;170(8):3375–3383. doi: 10.1128/jb.170.8.3375-3383.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Willey J., Santamaria R., Guijarro J., Geistlich M., Losick R. Extracellular complementation of a developmental mutation implicates a small sporulation protein in aerial mycelium formation by S. coelicolor. Cell. 1991 May 17;65(4):641–650. doi: 10.1016/0092-8674(91)90096-h. [DOI] [PubMed] [Google Scholar]
  49. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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