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. 1994 Sep;176(17):5530–5533. doi: 10.1128/jb.176.17.5530-5533.1994

Purification of the LysR family regulator, ClcR, and its interaction with the Pseudomonas putida clcABD chlorocatechol operon promoter.

W M Coco 1, M R Parsek 1, A M Chakrabarty 1
PMCID: PMC196742  PMID: 8071232

Abstract

Previous studies have shown that the clcABD operon is under the transcriptional control of the LysR-type activator ClcR. In this study, the conditions leading to its aggregation were avoided and ClcR was purified and confirmed by amino-terminal sequencing. Gel filtration indicated that ClcR exists as a dimer in solution. The DNase I footprint of ClcR was determined. The binding properties of ClcR and the catechol operon regulator, CatR, were compared.

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Selected References

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  1. Bouvet P., Belasco J. G. Control of RNase E-mediated RNA degradation by 5'-terminal base pairing in E. coli. Nature. 1992 Dec 3;360(6403):488–491. doi: 10.1038/360488a0. [DOI] [PubMed] [Google Scholar]
  2. Coco W. M., Rothmel R. K., Henikoff S., Chakrabarty A. M. Nucleotide sequence and initial functional characterization of the clcR gene encoding a LysR family activator of the clcABD chlorocatechol operon in Pseudomonas putida. J Bacteriol. 1993 Jan;175(2):417–427. doi: 10.1128/jb.175.2.417-427.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Englard S., Seifter S. Precipitation techniques. Methods Enzymol. 1990;182:285–300. doi: 10.1016/0076-6879(90)82024-v. [DOI] [PubMed] [Google Scholar]
  4. Goethals K., Van Montagu M., Holsters M. Conserved motifs in a divergent nod box of Azorhizobium caulinodans ORS571 reveal a common structure in promoters regulated by LysR-type proteins. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1646–1650. doi: 10.1073/pnas.89.5.1646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jaeger J. A., Turner D. H., Zuker M. Improved predictions of secondary structures for RNA. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7706–7710. doi: 10.1073/pnas.86.20.7706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jaeger J. A., Turner D. H., Zuker M. Predicting optimal and suboptimal secondary structure for RNA. Methods Enzymol. 1990;183:281–306. doi: 10.1016/0076-6879(90)83019-6. [DOI] [PubMed] [Google Scholar]
  7. Neidle E. L., Hartnett C., Ornston L. N. Characterization of Acinetobacter calcoaceticus catM, a repressor gene homologous in sequence to transcriptional activator genes. J Bacteriol. 1989 Oct;171(10):5410–5421. doi: 10.1128/jb.171.10.5410-5421.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Parsek M. R., Shinabarger D. L., Rothmel R. K., Chakrabarty A. M. Roles of CatR and cis,cis-muconate in activation of the catBC operon, which is involved in benzoate degradation in Pseudomonas putida. J Bacteriol. 1992 Dec;174(23):7798–7806. doi: 10.1128/jb.174.23.7798-7806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Parsek M. R., Ye R. W., Pun P., Chakrabarty A. M. Critical nucleotides in the interaction of a LysR-type regulator with its target promoter region. catBC promoter activation by CatR. J Biol Chem. 1994 Apr 15;269(15):11279–11284. [PubMed] [Google Scholar]
  10. Rothmel R. K., Shinabarger D. L., Parsek M. R., Aldrich T. L., Chakrabarty A. M. Functional analysis of the Pseudomonas putida regulatory protein CatR: transcriptional studies and determination of the CatR DNA-binding site by hydroxyl-radical footprinting. J Bacteriol. 1991 Aug;173(15):4717–4724. doi: 10.1128/jb.173.15.4717-4724.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Schell M. A. Molecular biology of the LysR family of transcriptional regulators. Annu Rev Microbiol. 1993;47:597–626. doi: 10.1146/annurev.mi.47.100193.003121. [DOI] [PubMed] [Google Scholar]
  12. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  14. Zuker M. On finding all suboptimal foldings of an RNA molecule. Science. 1989 Apr 7;244(4900):48–52. doi: 10.1126/science.2468181. [DOI] [PubMed] [Google Scholar]
  15. van der Meer J. R., Frijters A. C., Leveau J. H., Eggen R. I., Zehnder A. J., de Vos W. M. Characterization of the Pseudomonas sp. strain P51 gene tcbR, a LysR-type transcriptional activator of the tcbCDEF chlorocatechol oxidative operon, and analysis of the regulatory region. J Bacteriol. 1991 Jun;173(12):3700–3708. doi: 10.1128/jb.173.12.3700-3708.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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