Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Sep;170(9):3843–3846. doi: 10.1128/jb.170.9.3843-3846.1988

Identification of a celE-binding protein and its potential role in induction of the celE gene in Thermomonospora fusca.

E S Lin 1, D B Wilson 1
PMCID: PMC211379  PMID: 3410818

Abstract

Thermomonospora fusca cellulase E5 is encoded by the celE gene. This gene appears to be regulated at the transcriptional level by both induction and repression, and three putative closely linked promoters have been located by S1 mapping. To study its regulatory mechanism, a gel retardation assay was used to identify a protein in T. fusca cell extracts that interacted specifically with the DNA fragment containing the celE promoters. It was found that the binding activity appeared only when cellulase synthesis was induced, and it therefore resembled an activator protein involved in cellulase induction. DNase I footprinting identified the target sequence for this protein as a 21-base-pair sequence downstream from the putative celE promoters. The level of this protein was measured in two cellulase constitutive mutants, and the results suggest a complex control for celE induction.

Full text

PDF
3843

Images in this article

3844Image
on p.3844
3844Image
on p.3844
3845Image
on p.3845
3845Image
on p.3845

Selected References

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

  1. Frankel A. D., Ackers G. K., Smith H. O. Measurement of DNA-protein equilibria using gel chromatography: application to the HinfI restriction endonuclease. Biochemistry. 1985 Jun 4;24(12):3049–3054. doi: 10.1021/bi00333a037. [DOI] [PubMed] [Google Scholar]
  2. Geider K., Hoffmann-Berling H. Proteins controlling the helical structure of DNA. Annu Rev Biochem. 1981;50:233–260. doi: 10.1146/annurev.bi.50.070181.001313. [DOI] [PubMed] [Google Scholar]
  3. Hahn S., Hendrickson W., Schleif R. Transcription of Escherichia coli ara in vitro. The cyclic AMP receptor protein requirement for PBAD induction that depends on the presence and orientation of the araO2 site. J Mol Biol. 1986 Apr 5;188(3):355–367. doi: 10.1016/0022-2836(86)90160-9. [DOI] [PubMed] [Google Scholar]
  4. Irani M. H., Orosz L., Adhya S. A control element within a structural gene: the gal operon of Escherichia coli. Cell. 1983 Mar;32(3):783–788. doi: 10.1016/0092-8674(83)90064-8. [DOI] [PubMed] [Google Scholar]
  5. Lin E. S., Wilson D. B. Transcription of the celE gene in Thermomonospora fusca. J Bacteriol. 1988 Sep;170(9):3838–3842. doi: 10.1128/jb.170.9.3838-3842.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lin E., Wilson D. B. Regulation of beta-1,4-Endoglucanase Synthesis in Thermomonospora fusca. Appl Environ Microbiol. 1987 Jun;53(6):1352–1357. doi: 10.1128/aem.53.6.1352-1357.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  8. McClure W. R. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. doi: 10.1146/annurev.bi.54.070185.001131. [DOI] [PubMed] [Google Scholar]
  9. Pabo C. O., Sauer R. T. Protein-DNA recognition. Annu Rev Biochem. 1984;53:293–321. doi: 10.1146/annurev.bi.53.070184.001453. [DOI] [PubMed] [Google Scholar]
  10. Revzin A., von Hippel P. H. Direct measurement of association constants for the binding of Escherichia coli lac repressor to non-operator DNA. Biochemistry. 1977 Nov 1;16(22):4769–4776. doi: 10.1021/bi00641a002. [DOI] [PubMed] [Google Scholar]
  11. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  12. Rouvière-Yaniv J., Yaniv M., Germond J. E. E. coli DNA binding protein HU forms nucleosomelike structure with circular double-stranded DNA. Cell. 1979 Jun;17(2):265–274. doi: 10.1016/0092-8674(79)90152-1. [DOI] [PubMed] [Google Scholar]
  13. Sheflin L. G., Kowalski D. Altered DNA conformations detected by mung bean nuclease occur in promoter and terminator regions of supercoiled pBR322 DNA. Nucleic Acids Res. 1985 Sep 11;13(17):6137–6154. doi: 10.1093/nar/13.17.6137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tanaka I., Appelt K., Dijk J., White S. W., Wilson K. S. 3-A resolution structure of a protein with histone-like properties in prokaryotes. Nature. 1984 Aug 2;310(5976):376–381. doi: 10.1038/310376a0. [DOI] [PubMed] [Google Scholar]
  15. Winter R. B., von Hippel P. H. Diffusion-driven mechanisms of protein translocation on nucleic acids. 2. The Escherichia coli repressor--operator interaction: equilibrium measurements. Biochemistry. 1981 Nov 24;20(24):6948–6960. doi: 10.1021/bi00527a029. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES