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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
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28 references, click to expand
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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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