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PMID: 1900366 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Purification of the regulatory protein AlgR1 and its binding in the far upstream region of the algD promoter in Pseudomonas aeruginosa.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 5 ·1991-03-01 ·Pages 1760-4

Kato J, Chakrabarty AM

Abstract

A regulatory protein AlgR1, previously suggested to be a member of a two-component sensory transduction system because of its homology to OmpR and NtrC and its ability to allow activation of the algD promoter under conditions of high osmolarity, has been hyperproduced in Escherichia coli after deletion of the upstream region including part of the Shine-Dalgarno sequence of the algR1 gene and its subsequent cloning under the tac promoter. The AlgR1 protein is purified as a monomer, and the sequence of the nine N-terminal amino acids of the monomer matches with that predicted from the DNA sequence of the algR1 gene. The purified AlgR1 protein binds to two separate DNA fragments of the algD upstream region. DNase protection experiments identify these two DNA segments as 14-mer sequences centered at -382 and -458 regions, which contain a common CCGT-TCGTC sequence in them. While the presence of at least one AlgR1 binding site is important for the activation of the algD promoter, the presence of both of the binding sites in the upstream region leads to a higher level of activation.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,isolation & purification,metabolism Base Sequence Binding Sites DNA-Binding Proteins/genetics,isolation & purification,metabolism Deoxyribonuclease I Escherichia coli/genetics Gene Expression Regulation, Bacterial Molecular Sequence Data Nucleotide Mapping Plasmids Promoter Regions, Genetic Pseudomonas aeruginosa/genetics Recombinant Proteins/isolation & purification,metabolism Restriction Mapping
Chemicals
Bacterial Proteins DNA-Binding Proteins Recombinant Proteins AlgR1 protein, Pseudomonas syringae Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kato J
Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago 60612.
Chakrabarty A M
References (28)
28 references, click to expand
  1. Control of alginate synthesis in Pseudomonas aeruginosa: regulation of the algR1 gene.
    Biochem Biophys Res Commun. 1989 Oct 31;164(2):601-8 PMID: 2510718
  2. Protein phosphorylation and regulation of adaptive responses in bacteria.
    Microbiol Rev. 1989 Dec;53(4):450-90 PMID: 2556636
  3. High osmolarity is a signal for enhanced algD transcription in mucoid and nonmucoid Pseudomonas aeruginosa strains.
    J Bacteriol. 1989 May;171(5):2312-7 PMID: 2496102
  4. The algR gene, which regulates mucoidy in Pseudomonas aeruginosa, belongs to a class of environmentally responsive genes.
    J Bacteriol. 1989 Mar;171(3):1278-83 PMID: 2493441
  5. Identification of the DNA-binding domain of the OmpR protein required for transcriptional activation of the ompF and ompC genes of Escherichia coli by in vivo DNA footprinting.
    J Biol Chem. 1989 Jun 15;264(17):10104-9 PMID: 2656704
  6. Regulation of ribosomal RNA promoters with a synthetic lac operator.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):6929-33 PMID: 6390428
  7. Production of single-stranded plasmid DNA.
    Methods Enzymol. 1987;153:3-11 PMID: 3323803
  8. Purification and characterization of the OmpR protein, a positive regulator involved in osmoregulatory expression of the ompF and ompC genes in Escherichia coli.
    J Biol Chem. 1986 Nov 15;261(32):15252-6 PMID: 3533941
  9. A bacterial enhancer functions to tether a transcriptional activator near a promoter.
    Science. 1990 Apr 27;248(4954):486-90 PMID: 1970441
  10. Pulmonary dehydration and infection in cystic fibrosis: evidence that ethanol activates alginate gene expression and induction of mucoidy in Pseudomonas aeruginosa.
    Mol Microbiol. 1990 May;4(5):737-45 PMID: 2167423
  11. DNA-looping and enhancer activity: association between DNA-bound NtrC activator and RNA polymerase at the bacterial glnA promoter.
    Proc Natl Acad Sci U S A. 1990 Jul;87(14):5504-8 PMID: 2164685
  12. Activation of the Klebsiella pneumoniae nifU promoter: identification of multiple and overlapping upstream NifA binding sites.
    Nucleic Acids Res. 1990 Apr 11;18(7):1693-701 PMID: 2186362
  13. AlgR3, a protein resembling eukaryotic histone H1, regulates alginate synthesis in Pseudomonas aeruginosa.
    Proc Natl Acad Sci U S A. 1990 Apr;87(8):2887-91 PMID: 2109318
  14. DNA looping in cellular repression of transcription of the galactose operon.
    Genes Dev. 1990 Mar;4(3):410-8 PMID: 2186968
  15. A control element within a structural gene: the gal operon of Escherichia coli.
    Cell. 1983 Mar;32(3):783-8 PMID: 6299576
  16. Genetic analysis of the ompB locus in Escherichia coli K-12.
    J Mol Biol. 1981 Sep 5;151(1):1-15 PMID: 6799653
  17. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity.
    Nucleic Acids Res. 1978 Sep;5(9):3157-70 PMID: 212715
  18. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  19. A new method for sequencing DNA.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
  20. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  21. Role of the far-upstream sites of the algD promoter and the algR and rpoN genes in environmental modulation of mucoidy in Pseudomonas aeruginosa.
    J Bacteriol. 1990 Nov;172(11):6576-80 PMID: 2121718
  22. The influence of the Klebsiella pneumoniae regulatory gene nifL upon the transcriptional activator protein NifA.
    Mol Microbiol. 1990 Aug;4(8):1253-8 PMID: 2280685
  23. Transcription of glnA in E. coli is stimulated by activator bound to sites far from the promoter.
    Cell. 1986 Jun 20;45(6):785-92 PMID: 2871943
  24. Protein kinase and phosphoprotein phosphatase activities of nitrogen regulatory proteins NTRB and NTRC of enteric bacteria: roles of the conserved amino-terminal domain of NTRC.
    Proc Natl Acad Sci U S A. 1988 Jul;85(14):4976-80 PMID: 2839825
  25. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  26. Dual mechanism of repression at a distance in the lac operon.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):8968-72 PMID: 3143112
  27. Pseudomonas aeruginosa infection in cystic fibrosis: nucleotide sequence and transcriptional regulation of the algD gene.
    Nucleic Acids Res. 1987 Jun 11;15(11):4567-81 PMID: 3108855
  28. Nucleotide sequence of a regulatory region controlling alginate synthesis in Pseudomonas aeruginosa: characterization of the algR2 gene.
    Gene. 1989 Dec 7;84(1):31-8 PMID: 2514124
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.5.1760
Published
1991-03-01
Pages
1760-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51104
Subset
IM
Grants
NIAID NIH HHS · AI 16790-12 · United States
Databases
GENBANK
M58062, M58063, M58064, M58065, M58066, M58067, M58705, M59766, M59767, M59768
External Links
PubMed source DOI: 10.1073/pnas.88.5.1760 PMC Full Text View journal details
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