Abstract
In previous experiments, aStreptomyces aureofaciens gene highly similar to the sporulation-specificwhiB gene ofStreptomyces cœlicolor was identified. By intergrative transformationvia double cross-over, a stable null mutant of thewhiB-homologous gene ofS. aureofaciens was obtained. The disruption blocked differentiation at a stage between the formation of aerial mycelium and the development of mature spores, producing white aerial hyphae without septation. Expression of thewhiB gene was investigated during differentiation by S1 nuclease mapping, using RNA prepared fromS. aureofaciens in various developmental stages. Two putative promoters were identified upstream of thewhiB coding region. The stronger promoter,whiB-P2, was induced at the beginning of aerial mycelium formation, and the weaker promoter,whiB-P1, was expressed fairly constantly during differentiation. No differences in the expression of thewhiB promoters were detected in anrpoZ-disruptedS. aureofaciens strain. The promoter bearing DNA fragment was inserted into the promoter-probe vector pARC1 to produce an expression pattern consistent with the results of direct RNA analysis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ausubel F.M., Brent R., Kingston R.E., Moore D.O., Seidman J.S., Smith J.A., Struhl K.:Current Protocols in Molecular Biology, Wiley, New York 1987.
Chater K.F.: A morphological and genetic mapping study of white colony mutants ofStreptomyces cœlicolor.J. Gen. Microbiol.72, 9–28 (1972).
Chater K.F.: Genetics of differentiationiinStreptomyces.Ann. Rev. Microbiol.47, 685–713 (1993).
Davis N.K., Chater K.F.: TheStreptomyces cœlicolor whiB gene encodes a small transcription factor-like protein dispensable for growth but essential for sporulation.Mol. Gen. Genet.232, 351–358 (1992).
Chater K.F., Merrick M.J.: Streptomycetes, pp. 93–114 in J.H. Parish (Ed.):Developmental Biology of Prokaryotes. Blackwell Scientific Publications, Oxford 1979.
Hopwood D.A., Bibb M.J., Chater K.F., Kieser T., Bruton C.J., Kieser H.M., Lydiate D.L., Smith C.P., Ward J.M., Schrempf H.:Genetic Manipulation of Streptomyces. A Laboratory Manual. The John Innes Foundation, Norwich 1985.
Hopwood D.A., Kieser T., Wright H.M., Biss M.J.: Plasmids, recombination, and chromosome mapping inStreptomyces lividans 66.J. Gen. Microbiol.129, 2257–2269 (1983).
Horinouchi S., Beppu T.: Construction and application of a promoter-probe plasmid that allows chromogenic identification inStreptomyces lividans.J. Bacteriol.162, 406–412 (1985).
Horinouchi S., Hara O., Beppu T.: Cloning of a pleiotropic gene that positively controls biosynthesis of A-factor, actinorhodin, and prodigiosin inStreptomyces cœlicolor A3(2) andStreptomyces lividans.J. Bacteriol.155, 1238–1248 (1983).
King E.J., Brown M.F.: A technique for preserving aerial fungal structure for scanning electron microscopy.Can. J. Microbiol.29, 653–658 (1983).
Kormanec J., Farkašovský M.: Differential expression of principal σ factor homologues ofStreptomyces aureofaciens correlates with the developmental stage.Nucl. Acids Res.21, 3647–3652 (1993).
Kormanec J., Farkašovský M.: Isolation of total RNA from yeast and bacteria and detection of rRNA in Northern blots.Bio-Techniques17, 838–842 (1994).
Kormanec J., Homerova D.:Streptomyces aureofaciens whiB gene encoding putative transcription factor essential for differentiation.Nucl. Acids Res.21, 2512 (1993).
Kormanec J., Potúčková L., Řežuchová B.: TheStreptomyces aureofaciens homologue of thewhiG encoding a putative σ factor essential for sporulation.Gene143, 101–103 (1994).
Kormanec J., Řežuchová B., Farkašovský M.: Optimization ofStreptomyces aureofaciens transformation and disruption of thehrdA gene encoding a homologue of the principal σ factor.J. Gen. Microbiol.139, 2525–2529 (1993).
Maxam A.M., Gilbert W.: Sequencing end-labelled DNA with base specific chemical cleavages.Methods Enzymol.65, 449–560 (1980).
Merrick M.J.: A morphological and genetic mapping study of bald colony mutants ofStreptomyces cœlicolor.J. Gen. Microbiol.96, 299–315 (1976).
Muth G., Nussbaumer B., Wohlleben W., Puhler A.: A vector system with temperature-sensitive replication for gene disruption and mutational cloning in streptomycetes.Mol. Gen. Genet.219, 341–348 (1989).
Soliveri J., Brown K.L., Buttner M.J., Chater K.F.: Two promoters for thewhiB sporulation gene ofStreptomyces cœlicolor A3(2) and their activities in relation to development.J. Bacteriol.174, 6215–6220 (1992).
Strohl W.R.: Compilation and analysis of DNA sequences associated with apparent streptomycete promoters.Nucl. Acids Res.20, 961–974 (1992).
Ward J.M., Jansen G.R., Kieser T., Buttner M.J., Bibb M.J.: Construction and characterization of a series of multi-copy promoter-probe plasmid vectors forStreptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicator.Mol. Gen. Genet.203, 468–478 (1986).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kormanec, J., Ševčíková, B., Sprušanský, O. et al. TheStreptomyces aureofaciens homologue of thewhiB gene is essential for sporulation; Its expression correlates with the developmental stage. Folia Microbiol 43, 605–612 (1998). https://doi.org/10.1007/BF02816376
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF02816376