1887

Microbiology Society logo

Volume 143, Issue 3

Activation of the subsp. pectin lyase structural gene : a role for RdgB Free

Abstract

The activation of pectin lyase (Pnl) production in subsp. strain 71 occurs upon DNA damage via a unique regulatory circuit involving and In a similar Pnl-inducible system reconstituted in , the product was found to activate the expression of , the structural gene for pectin lyase. The kinetic data presented here also show that transcription of followed that of in subsp. , indicating a temporal order of gene expression. By deletion analysis we have localized the promoter/regulatory region within a 66 bp DNA segment upstream of the transcriptional start site. This region contains the -10 consensus sequence but not the sequences corresponding to the -35 region. For DNA-binding studies, was overexpressed in and a 14 kDa polypeptide was identified as the gene product. RdgB from crude extracts or a purified preparation caused an identical gel mobility shift of a 164 bp DNA segment containing the promoter/regulatory region. Utilizing DNase I protection assay the RdgB-binding site was localized between nucleotides -29 and -56, i.e. overlapping the position of the putative -35 box. The findings reported here, taken along with our previous observation that the product is required for expression, establishes that encodes a transcriptional factor which specifically interacts with the promoter/regulatory region.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): cell-wall-degrading enzyme , damage-inducible traits , gene expression , plant-pathogenic bacteria and virulence factor
Society for General Microbiology 1997
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-3-705
1997-03-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/3/mic-143-3-705.html?itemId=/content/journal/micro/10.1099/00221287-143-3-705&mimeType=html&fmt=ahah

References

  1. Aiba H., Adhya S., Crombrugghe B. 1981; Evidence for two functional gal promoters in intact Escherichia coli. . J Biol Chem 256:11905–11910
     [Google Scholar]
  2. Barras F., van Gijsegem F., Chatterjee A.K. 1994; Extracellular enzymes and pathogenesis of soft-rot Erwinia. . Annu Rev Phytopathol 32:201–234
     [Google Scholar]
  3. Bölker M., Wulczyn F.G., Kahmann R. 1989; Role of bacteriophage Mu C protein in activation of the mom gene promoter.. J Bacteriol 171:2019–2027
     [Google Scholar]
  4. Brosius J. 1984; Plasmid vectors for the selection of promoters.. Gene 27:151–160
     [Google Scholar]
  5. Chatterjee A., McEvoy J.L., Chambost J.P., Blasco F., Chatterjee A.K. 1991; Nucleotide sequence and molecular characterization of pnlA, the structural gene for damage- inducible pectin lyase of Erwinia carotovora subsp.carotovora 71.. J Bacteriol 173:1765–1769
     [Google Scholar]
  6. Harley C.B., Reynolds R.P. 1987; Analysis of E. coli promoter sequences.. Nucleic Acids Res 15:2343–2361
     [Google Scholar]
  7. Ho Y.S., Wulff D.L., Rosenberg R.P. 1983; Bacteriophage λprotein dl binds promoters on the opposite face of the DNA helix from RNA polymerase.. Nature 304:703–708
     [Google Scholar]
  8. Hochschild A., Irwin N. 1983; Repressor structure and the mechanism of positive control.. Cell 32:319–325
     [Google Scholar]
  9. Itoh Y., Izaki K., Takahashi H. 1980; Simultaneous synthesis of pectin lyase and carotovoricin induced by mitomycin C, nalidixic acid or ultraviolet light irradiation in Erwinia carotovora. . Agric Biol Chem 44:1135–1140
     [Google Scholar]
  10. Kamimiya S., Izaki K., Takahashi H. 1972; A new pectolytic enzyme in Erwinia aroideae formed in the presence of nalidixic acid.. Agric Biol Chem 36:2367–2372
     [Google Scholar]
  11. Kurland C.G., Dong H. 1996; Bacterial growth inhibition by overproduction of protein.. Mol Microbiol 21:1–4
     [Google Scholar]
  12. Leblanc B., Moss T. 1994; DNase I footprinting.. In DNA- Protein Interaction: Principles and Protocols pp. 1–10 Kneale G.G. Edited by Totowa, NJ: Humana Press;
     [Google Scholar]
  13. Lee N.L., Gielow W.O., Wallace R.G. 1981; Mechanism of araC autoregulation and the domains of two overlapping promoters, Pc and PBAD, in the L-arabinose regulatory region of Escherichia coli. . Proc Natl Acad Sci USA 78752–756
     [Google Scholar]
  14. Liu Y., Chatterjee A.K. 1996; Analysis of the RecA-Rdg regulatory circuit that controls pectin lyase production in Erwinia carotovora subsp.carotovora (Ecc): evidence for repression of rdgA and rdgB by RdgA.. In Abstracts of the 8th International Congress on Molecular Plant-Microbe Interactions Knoxville, TN, USA: (Abstracts), G-10
    [Google Scholar]
  15. Liu Y., Chatterjee A., Chatterjee A.K. 1994; Nucleotide sequence, organization and expression of rdgA and rdgB genes that regulate pectin lyase production in the plant pathogenic bacterium Erwinia carotovora subsp.carotovora in response to DNA-damaging agents.. Mol Microbiol 14:999–1010
     [Google Scholar]
  16. Liu Y., Wang X., Mukherjee A., Chatterjee A.K. 1996; RecA relieves negative autoregulation of rdgA which specifies a component of the RecA-Rdg regulatory circuit controlling pectin lyase production in Erwinia carotovora subsp.carotovora. . Mol Microbiol 22:909–918
     [Google Scholar]
  17. McEvoy J.L., Thurn K.K., Chatterjee A.K. 1987; Expression of the E. coli lexA+ gene in Erwinia carotovora subsp.carotovora and its effect on production of pectin lyase and carotovoricin.. FEMS Microbiol Lett 42:205–208
     [Google Scholar]
  18. McEvoy J.L., Murata H., Chatterjee A.K. 1990; Molecular cloning and characterization of an Erwinia carotovora subsp.carotovora pectin lyase gene that responds to DNA-damaging agents.. J Bacteriol 172:3284–3289
     [Google Scholar]
  19. McEvoy J.L., Murata H., Chatterjee A.K. 1992; Genetic evidence for an activator required for induction of pectin lyase in Erwinia carotovora subsp.carotovora by DNA-damaging agents.. J Bacteriol 174:5471–5474
     [Google Scholar]
  20. Margolin W., Rao G., Howe M.M. 1989; Bacteriophage Mu late promoters: four late transcripts initiate near a conserved sequence.. J Bacteriol 171:2003–2018
     [Google Scholar]
  21. Pirhonen M., Saarilahti H., Karlsson M.-B., Palva E.T. 1991; Identification of pathogenicity determinants of Erwinia carotovora subsp.carotovora by transposon mutagenesis.. Mol Plant-Microbe Interact 4:276–283
     [Google Scholar]
  22. Raibaud O., Schwartz M. 1984; Positive control of transcription initiation in bacteria.. Annu Rev Genet 18:173–206
     [Google Scholar]
  23. Reverchon S., Huang Y., Bourson C., Robert-Baudouy J. 1989; Nucleotide sequences of Erwinia chrysantbemi ogl and pelE genes negatively regulated by the kdgR product.. Gene 85:125–134
     [Google Scholar]
  24. Reznikoff W.S., McClure W.R. 1986; E. coli promoters.. In Maximizing Gene Expression pp. 1–33 Reznikoff W.S., Gold L. Edited by Boston: Butterworths;
     [Google Scholar]
  25. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  26. Shaw W.V. 1975; Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria.. Methods Enzymol 43:737–755
     [Google Scholar]
  27. Tabor S., Richardson C.C. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.. Proc Natl Acad Sci USA 821074–1078
     [Google Scholar]
  28. Tsuyumu S., Chatterjee A.K. 1984; Pectin lyase production in Erwinia chrysantbemi and other soft-rot Erwinia species.. Physiol Plant Pathol 24:291–302
     [Google Scholar]
  29. Tomizawa H., Takahashi H. 1971; Stimulation of pectolytic enzyme formation of Erwinia aroideae by nalidixic acid, mitomycin C and bleomycin.. Agric Biol Chem 35:191–200
     [Google Scholar]
  30. Tsuyumu S., Funakubo T., Chatterjee A.K. 1985; Purification and partial characterization of a mitomycin C-induced pectin lyase of Erwinia chrysantbemi strain EC183.. Physiol Plant Pathol 27:119–130
     [Google Scholar]
  31. Zink R.T., Kemble R.J., Chatterjee A.K. 1984; Transposon Tn5 mutagenesis in Erwinia carotovora subsp.carotovora and E. carotovora subsp.atroseptica. . J Bacteriol 157:809–814
     [Google Scholar]
  32. Zink R.T., Engwall J.K., McEvoy J.L., Chatterjee A.K. 1985; recA is required in the induction of pectin lyase and carotovoricin in Erwinia carotovora subsp.carotovora. . J Bacteriol 164:390–396
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-3-705
Loading
Activation of the Erwinia carotovora subsp. carotovora pectin lyase structural gene pnIA: a role for RdgB
Microbiology 143, 705 (1997); https://doi.org/10.1099/00221287-143-3-705
/content/journal/micro/10.1099/00221287-143-3-705
/content/journal/micro/10.1099/00221287-143-3-705
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error