1887

Abstract

Expression of the gene encoding a sporulation-specific sigma factor, σ, in is restricted only to sporulation. Gel mobility-shift assays using protein fractions from different developmental stages of revealed two different putative proteins specifically bound to the promoter region: a protein (designated RsfA) present in young substrate mycelium, and a protein (designated RsfB) present in the course of sporulation. Based on the characteristic profiles of their appearance during differentiation, RsfA might be a repressor and RsfB an activator of expression. The location of a specific binding site of the repressor-like protein (RsfA) was determined by gel mobility-shift assays of promoter deletion fragments and by DNase I footprinting analysis. The binding site mapped from nucleotides −87 to −25 relative to the transcription start point of the promoter, and overlapped the −35 promoter region. Given the dependence of expression upon , the putative sporulation transcription factor WhiH was overproduced in and used in the mobility-shift assays with the promoter. However, no specific binding was detected, indicating an indirect dependence of upon .

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2000-11-01
2020-04-04
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References

  1. Aı́nsa J. A., Parry H. D., Chater K. F.. 1999; A response regulator-like protein that functions at an intermediate stage of sporulation in Streptomyces coelicolor A3(2). Mol Microbiol34:607–619[CrossRef]
    [Google Scholar]
  2. Ausubel F. M., Brent R., Kingston R. E., Moore D. O., Seidman J. S., Smith J. A., Struhl K.. 1987; Current Protocols in Molecular Biology New York: Wiley;
    [Google Scholar]
  3. Bradford M. M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem72:248–254[CrossRef]
    [Google Scholar]
  4. Chater K. F.. 1998; Taking a genetic scalpel to the Streptomyces colony. Microbiology144:1465–1478[CrossRef]
    [Google Scholar]
  5. Chater K. F., Bruton C. J., Plaskitt K. A., Buttner M. J., Mendez C., Helmann J. D.. 1989; The developmental fate of S. coelicolor hyphae depends upon a gene product homologous with the motility σ factor of B. subtilis. Cell59:133–143[CrossRef]
    [Google Scholar]
  6. Davis N. K., Chater K. F.. 1992; The Streptomyces coelicolor whiB gene encodes a small transcription factor-like protein dispensable for growth but essential for sporulation. Mol Gen Genet232:351–358
    [Google Scholar]
  7. Gralla J. D.. 1990; Promoter recognition and mRNA initiation by Escherichia coli70. Methods Enzymol185:37–54
    [Google Scholar]
  8. Hopwood D. A., Bibb M. J., Chater K. F..7 other authors 1985; Genetic Manipulation of Streptomyces: a Laboratory Manual Norwich: The John Innes Foundation;
    [Google Scholar]
  9. Horinouchi S., Hara O., Beppu T.. 1983; Cloning of a pleiotropic gene that positively controls biosynthesis of A-factor, actinorhodin, and prodigiosin in Streptomyces coelicolor A3(2) and Streptomyces lividans. J Bacteriol155:1238–1248
    [Google Scholar]
  10. Kelemen G. H., Brown G. L., Kormanec J., Potúčková L., Chater K. F., Buttner M. J.. 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 Microbiol21:593–603[CrossRef]
    [Google Scholar]
  11. Kormanec J., Farkašovský M.. 1993; Differential expression of principal sigma factor homologues of Streptomyces aureofaciens correlates with developmental stage. Nucleic Acids Res21:3647–3652[CrossRef]
    [Google Scholar]
  12. Kormanec J., Potúčková L., Řežuchová B.. 1994; The Streptomyces aureofaciens homologue of the whiG gene encoding a putative sigma factor essential for sporulation. Gene143:101–103[CrossRef]
    [Google Scholar]
  13. Kormanec J., Homerová D., Potúčková L., Nováková R., Řežuchová B.. 1996; Differential expression of two sporulation specific σ factors of Streptomyces aureofaciens correlates with the developmental stage. Gene181:19–27[CrossRef]
    [Google Scholar]
  14. Kormanec J., Ševčı́ková B., Sprušanský O., Benada O., Kofroňová O., Novákova� R., Řežuchová B., Potúčková L., Homérová D.. 1998; The Streptomyces aureofaciens homologue of the whiB gene is essential for sporulation and its expression correlates with the developmental stage. Folia Microbiol43:605–612[CrossRef]
    [Google Scholar]
  15. Kormanec J., Nováková R., Homerová D., Ševčı́ková B.. 1999; The Streptomyces aureofaciens homologue of the sporulation gene whiH is dependent on rpoZ-encoded σ factor. Biochim Biophys Acta1444:80–84[CrossRef]
    [Google Scholar]
  16. Kroos L., Zhang B., Ichikawa H., Yu Y.-T. N.. 1999; Control of σ factor activity during Bacillus subtilis sporulation. Mol Microbiol31:1285–1294[CrossRef]
    [Google Scholar]
  17. Laemmli U. K.. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature227:680–685[CrossRef]
    [Google Scholar]
  18. Maxam A. M., Gilbert W.. 1980; Sequencing end-labelled DNA with base specific chemical cleavages. Methods Enzymol65:449–560
    [Google Scholar]
  19. Potúčková L., Kelemen G. H., Findlay K. C., Lonetto M. A., Buttner M. J., Kormanec J.. 1995; A new RNA polymerase sigma factor, σF, is required for the late stages of morphological differentiation in Streptomyces spp. Mol Microbiol17:37–48[CrossRef]
    [Google Scholar]
  20. Řežuchová B., Barák I., Kormanec J.. 1997; Disruption of a sigma factor gene, sigF, affects an intermediate stage of spore pigment production in Streptomyces aureofaciens. FEMS Microbiol Lett153:371–377[CrossRef]
    [Google Scholar]
  21. Ryding N. J., Kelemen G. H., Whatling C. A., Flardh K., Buttner M. J.. 1998; A developmentally regulated gene encoding a repressor-like protein is essential for sporulation in Streptomyces coelicolor A3(2). Mol Microbiol29:343–357[CrossRef]
    [Google Scholar]
  22. Strohl W. R.. 1992; Compilation and analysis of DNA sequences associated with apparent streptomycete promoters. Nucleic Acids Res20:961–974[CrossRef]
    [Google Scholar]
  23. Sun J., Kelemen G. H., Fernandez-Abalos J. M., Bibb M. J.. 1999; Green fluorescent protein as a reporter for spatial and temporal gene expression in Streptomyces coelicolor A3(2). Microbiology145:2221–2227
    [Google Scholar]
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