[en] The gram-positive filamentous bacterium Streptomyces coelicolor has a complex developmental cycle with three distinct phases: growth of the substrate mycelium, development of reproductive structures called aerial hyphae, and differentiation of these aerial filaments into long chains of exospores. During a transposon mutagenesis screen, we identified a novel gene (devA) required for proper development. The devA mutant produced only rare aerial hyphae, and those that were produced developed aberrant spore chains that were much shorter than wild-type chains and had misplaced septa. devA encodes a member of the GntR superfamily, a class of transcriptional regulators that typically respond to metabolite effector molecules. devA forms an operon with the downstream gene devB, which encodes a putative hydrolase that is also required for aerial mycelium formation on R5 medium. S1 nuclease protection analysis showed that transcription from the single devA promoter was temporally associated with vegetative growth, and enhanced green fluorescent protein transcriptional fusions showed that transcription was spatially confined to the substrate hyphae in the wild type. In contrast, devAB transcript levels were dramatically upregulated in a devA mutant and the devA promoter was also active in aerial hyphae and spores in this background, suggesting that DevA might negatively regulate its own production. This suggestion was confirmed by gel mobility shift assays that showed that DevA binds its own promoter region in vitro.
Disciplines :
Microbiology
Author, co-author :
Hoskisson, P. A.
Rigali, Sébastien ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Fowler, K.
Findlay, K. C.
Buttner, M. J.
Language :
English
Title :
DevA, a GntR-like transcriptional regulator required for development in Streptomyces coelicolor
Publication date :
July 2006
Journal title :
Journal of Bacteriology
ISSN :
0021-9193
eISSN :
1098-5530
Publisher :
Amer Soc Microbiology, Washington, United States - Washington
Aínsa, J. A., H. D. Parry, and K. F. Chafer. 1999. A response regulator-like protein that functions at an intermediate stage of sporulation in Streptomyces coelicolor A3(2). Mol. Microbiol. 34:607-619.
Bateman, A., E. Birney, L. Cerruti, R. Durbin, L. Etwiller, S. R. Eddy, S. Griffiths-Jones, K. L. Howe, M. Marshall, and E. L. Sonnhammer. 2002. The Pfam protein families database. Nucleic Acids Res. 30:276-280.
Bentley, S. D., K. F. Chafer, A-M. Cerdeño-Tárraga, G. L. Challis, N. R. Thomson, K. D. James, D. E. Harris, M. A. Quail, H. Kieser, D. Harper, A. Bateman, S. Brown, G. Chandra, C. W. Chen, M. Collins, A. Cronin, A. Fraser, A. Goble, J. Hidalgo, T. Hornshy, S. Howarth, C.-H. Huang, T. Kieser, L. Larke, L. Murphy, K. Oliver, S. O'Neil, E. Rabbinowitsch, M.-H. Rajandream, K. Rutherford, S. Rutter, K. Seeger, D. Saunders, S. Sbarp, R. Squares, S. Squares, K. Taylor, T. Warren, A. Wietzorrek, J. Woodward, B. G. Barrell, J. Parkhill, and D. A. Hopwood. 2002. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417:141-147.
Bihh, M. J., V. Molle, and M. J. Buttner. 2000. σBRIN, an extracytoplasmic function RNA polymerase sigma factor required for aerial mycelium formation in Streptomyces coelicolor A3(2). J. Bacteriol. 182:4606-4616.
Bierman, M., R. Logan, K. O'Brien, E. T. Seno, R. N. Rao, and B. E. Schoner. 1992. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43-49.
Bishop, A., S. Fielding, P. J. Dyson, and P. R. Herron. 2003. Concerted mutagenesis of a streptomyccte genome: a link between osmoadaptation, development and antibiotic production. Genome Res. 14:893-900.
Chater, K. F. 2001. Regulation of sporulation in Streptomyces coelicolor A3(2): a Checkpoint multiplex? Curr. Opin. Microbiol. 4:667-673.
Chater, K. F., and S. Horinouchi. 2003. Signalling early developmental events in two highly diverged Streptomyces species. Mol. Microbiol. 48:9-15.
Chung, S.-T. 1987. Tn4556, a 6.8-kilobase-pair transposable element of Streptomyces fradiae. J. Bacteriol. 169:4436-4441.
Dodd, I. B., and J. B. Egan. 1990. Improved detection of helix-turn-helix DNA-binding motifs in protein sequences. Nucleic Acids Res. 18:5019-5026.
DiRusso, C. C., and T. Nyström. 1998. The fats of Escherichia coli during infancy and old age: regulation by global regulators, alarmones and lipid intermediates. Mol. Microbiol. 27:1-8.
DiRusso, C. C., A. K. Metzger, and T. L. Heimert. 1993. Regulation of transcription of genes required for fatty acid transport and unsaturated fatty acid biosynthesis in Escherichia coli by FadR. Mol. Microbiol. 7:311-322.
DiRusso, C. C., T. L. Heimert, and A. K. Metzger. 1992. Characterization of FadR, a global transcriptional regulator of fatty acid metabolism in Escherichia coli. Interaction with the fadB promoter is prevented by long chain fatty acyl coenzyme A. J. Biol. Chcm. 267:8685-8691.
Elliot, M. A., and N. J. Talhot. 2004. Building filaments in the air: aerial morphogenesis in bacteria and fungi. Curr. Opin. Microbiol. 7:594-601.
Elliot, M. A., M. J. Bihb, M. J. Buttner, and B. K. Leskiw. 2001. BldD is a direct regulator of key developmental genes in Streptomyces coelicolor A3(2). Mol. Microbiol. 40:257-269.
Fowler, K. 2002. Transposon mutagenesis of Streptomyces coelicolor A3(2). Ph.D thesis. University of East Anglia. Norwich, United Kingdom.
Fujita, Y., and T. Fujita. 1987. The gluconate operon gut of Bacillus subtilis encodes its own transcriptional negative regulator. Proc. Natl. Acad. Sci. USA 84:4524-4528.
Fujita, Y., and Y. Miwa. 1989. Identification of an operator sequence for the Bacillus subtilis gut operon. J. Biol. Chem. 264:4201-4206.
Gehring, A. M., J. R. Nodwell, S. M. Beverley, and R. Losick. 2000. Genome-wide insertional mutagenesis in Streptomyces coelicolor reveals additional genes involved in morphological differentiation. Proc. Natl. Acad. Sci. USA 97:9642-9647.
Gehring, A. M., N. J. Yoo, and R. Losick. 2001. An RNA polymerase sigma factor that blocks morphological differentiation by Streptomyces coelicolor A3(2). J. Bacteriol. 183:5991-5996.
Gehring, A. M., S. T. Wang, D. B. Kearns, N. Y. Storer, and R. Losick. 2004. Novel genes that influence development in Streptomyces coelicolor. J. Bacteriol. 186:3570-3577.
Gregory, M. A., R. Till, and M. C. M. Smith. 2003. Integration site for Streptomyces phage φBTI and development of site-specific integrating vectors. J. Bacteriol 185:5320-5323.
Gust, B., G. L. Challis, K. Fowler, T. Kieser, and K. F. Chater. 2003. Gene replacement by PCR targeting in Streptomyces and its use to identify a protein domain involved in the biosynthesis of the sesquiterpene odour geosmin. Proc. Natl. Acad. Sci. USA 100:1541-1546.
Gust, B., G. Chandra, D. Jakimowicz, Y. Tian, C. J. Bruton, and K. F. Chater. 2004. λ Red-mediated genetic manipulation of antibiotic-producing Streptomyces. Adv. Appl. Microbiol. 54:107-128.
Haydon, D. J., and J. R. Guest. 1991. A new family of bacterial regulatory proteins. FEMS Microbiol. Lett. 63:291-295.
Henry, M. F., and J. E. Cronan. 1991. Escherichia coli transcription factor that both activates fatty acid synthesis and represses fatty acid degradation. J. Mol. Biol. 222:843-849.
Henry, M. F., and J. E. Cronan. 1992. A new mechanism of transcriptional regulation: release of an activator triggered by small molecule binding. Cell 70:671-679.
Janssen, G. R. 1993. Eubacterial, archaebactcrial and Eukaryotic genes that encode leaderless mRNA, p. 59-67. In R. H. Baltz, G. D. Hegeman, and P. L. Skatrud (ed.). Industrial microorganisms: basic and applied molecular genetics. American Society for Microbiology. Washington, D.C.
Karandikar, A., G. P. Sharples, and G. Hobbs. 1997. Differentiation of Streptomyces coelicolor A3(2) under nitrate-limited conditions. Microbiology 143:3581-3590.
Kelemen, G. H., and M. J. Buttner. 1998. Initiation of aerial mycelium formation in Streptomyces. Curr. Opin. Microbiol. 1:656-662.
Kelemen, G. H., G. L. Brown, J. Kormanec, L. Potûcková, K. F. Chater, and M. J. Buttner. 1996. The positions of the sigma-factor genes, whiG and sigF, in the hierarchy controlling the development of spore chains in the aerial hyphae of Streptomyces, coelicolor A3(2). Mol. Microbiol. 21:593-603.
Kieser, T., M. J. Bibb, M. J. Buttner, K. K. Chater, and D. A. Hopwood. 2000. Practical Streptomyces Genetics. John Innes Foundation, Norwich, United Kingdom.
Kodani, S., M. E. Hudson, M. C. Durrant, M. J. Buttner, J. R. Nodwell, and J. M. Willey. 2004. The SapB morphogen is a lantibiotic-like peptide derived from the product of the developmental gene ramS in Streptomyces coelicolor. Proc. Natl. Acad. Sci. USA 101:11448-11453.
Lee, M. H., M. Scherer, S. Rigali, and J. W. Golden. 2003. PlmA, a new member of the GntR family, has plasmid maintenance functions in Anabaena sp. strain PCC 7120. J. Bacteriol. 185:4315-4325.
Molle, V., W. J. Palframan, K. C. Findlay, and M. J. Buttner. 2000. WhiD and WhiB, homologous proteins required for different stages of sporulation in Streptomyces coelicolor A3(2). J. Bacteriol. 182:1286-1295.
Mota, L. J., P. Tavares, and I. Sá-Nogueira. 1999. Mode of action of AraR, the key regulator of L-arabinose metabolism in Bacillus subtilis. Mol. Microbiol. 33:476-489.
Nodwell, J. R., K. McGovern, and R. Losick. 1996. An oligopeptide permease responsible for the import of an extracellular signal governing aerial mycelium formation in Streptomyces coelicolor. Mol. Microbiol. 22:881-893.
Nodwell, J. R, M. Yang, D. Kuo, and R. Losick. 1999. Extracellular complementation and the identification of additional genes involved in aerial mycelium formation in Streptomyces coelicolor. Genetics 151:569-584.
O'Connor, T. J., P. Kanellis, and J. R. Nodwell. 2002. The ramC gene is required for morphogenesis in Streptomyces coelicolor and expressed in a cell type-specific manner under the direct control of RamR. Mol. Microbiol. 45:45-57.
Paget, M. S. B., L. Chamberlin, A. Atrih, S. J. Foster, and M. J. Buttner. 1999. Evidence that the extracytoplasmic function sigma factor, σE, is required for normal cell wall structure in Streptomyces coelicolor A3(2). J. Bacteriol. 181:204-211.
Pope, M. K., B. D. Green, and J. Westpheling. 1996. The bld mutants of Streptomyces coelicolor are defective in the regulation of carbon utilization, morphogenesis and cell-cell signalling. Mol. Microbiol. 19:747-756.
Quail, M. A., C. E. Dempsey, and J. R. Guest. 1994. Identification of a fatty acyl responsive regulator (FarR) in Escherichia coli. FEBS Lett. 356:183-187.
Quail, M. A., D. J. Haydon, and J. R. Guest. 1994. The pdhR-aceEF-lpd operon of Escherichia coli expresses the pyruvate dehydrogenase complex. Mol. Microbiol. 12:95-104.
Rigali, S., A. Derouaux, F. Giannotta, and J. Dusart. 2002. Subdivision of the helix-turn-helix GntR family of bacterial regulators in the FadR, HutC, MocR, and YtrA subfamilies. J. Biol. Chem. 277:12507-12515.
Rigali, S., M. Schlicht, P. A. Hoskisson, H. Northaft, M. Merzbacher, B. Joris, and F. Titgemeyer. 2004. Extending the classification of bacterial transcription factors beyond the helix-turn-helix motif as an alternative approach to discover new cis/trans relationships. Nucleic Acids Res. 32:3418-3426.
Ryding, N. J., G. H. Kelemen, C. A. Whatling, K. Flärdh, M. J. Buttner, and K. F. Chater. 1998. A developmentally regulated gene encoding a repressor-like protein is essential for sporulation in Streptomyces coelicolor A3(2). Mol. Microbiol. 29:343-357.
Ryding, N. J., M. J. Bibb, V. Molle, K. C. Findlay, K. F. Chater, and M. J. Buttner. 1999. New sporulation loci in Streptomyces coelicolor A3(2). J. Bacteriol. 181:5419-5425.
Seo, J. W., Y. Ohnishi, A. Hirata, and S. Horinouchi. 2002. ATP-binding cassette transport system involved in regulation of morphological differentiation in response to glucose in Streptomyces griseus. J. Bacteriol. 184:91-103.
Sprusansky, O., L. Zhou, S. Jordan, J. White, and J. Westpheling. 2003. Identification of three new genes involved in morphogenesis and antibiotic production in Streptomyces coelicolor. J. Bacteriol. 185:6147-6157.
Sun, J., G. H. Kelemen, J. M. Fernandez-Ahalos, and M. J. Bibb. 1999. Green fluorescent protein as a reporter for spatial and temporal gene expression in Streptomyces coelicolor A3(2). Microbiology 145:2221-2227.
Willey, J., J. Schwedock, and R. Losick. 1993. Multiple extracellular signals govern the production of a morphogenetic protein involved in aerial mycelium formation by Streptomyces coelicolor. Genes Dev. 7:895-903.