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PMID: 8824606 Published · ppublish English Comparative Study Journal Article

The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product.

Journal of bacteriology ·Vol. 178 ·No. 19 ·1996-10-00 ·Pages 5627-35

MacGregor CH, Arora SK, Hager PW, Dail MB, Phibbs PV

Abstract

The gene (crc) responsible for catabolite repression control in Pseudomonas aeruginosa has been cloned and sequenced. Flanking the crc gene are genes encoding orotate phosphoribosyl transferase (pyrE) and RNase PH (rph). New crc mutants were constructed by disruption of the wild-type crc gene. The crc gene encodes an open reading frame of 259 amino acids with homology to the apurinic/apyrimidinic endonuclease family of DNA repair enzymes. However, crc mutants do not have a DNA repair phenotype, nor can the crc gene complement Escherichia coli DNA repair-deficient strains. The crc gene product was overexpressed in both P. aeruginosa and in E. coli, and the Crc protein was purified from both. The purified Crc proteins show neither apurinic/apyrimidinic endonuclease nor exonuclease activity. Antibody to the purified Crc protein reacted with proteins of similar size in crude extracts from Pseudomonas putida and Pseudomonas fluorescens, suggesting a common mechanism of catabolite repression in these three species.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Cross Reactions DNA-(Apurinic or Apyrimidinic Site) Lyase Deoxyribonuclease IV (Phage T4-Induced) Enzyme Repression Escherichia coli/genetics Escherichia coli Proteins Exoribonucleases/genetics Genes, Bacterial Lyases/genetics Molecular Sequence Data Orotate Phosphoribosyltransferase/genetics Pseudomonas aeruginosa/genetics Recombinant Proteins/isolation & purification Repressor Proteins/genetics,immunology,isolation & purification Sequence Analysis, DNA Sequence Homology, Amino Acid Species Specificity
Chemicals
Bacterial Proteins Escherichia coli Proteins Recombinant Proteins Repressor Proteins crc protein, Pseudomonas Orotate Phosphoribosyltransferase ribonuclease PH Exoribonucleases Deoxyribonuclease IV (Phage T4-Induced) endonuclease IV, E coli Lyases DNA-(Apurinic or Apyrimidinic Site) Lyase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
MacGregor C H
Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina 27858, USA.
Arora S K
Hager P W
Dail M B
Phibbs P V
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.178.19.5627-5635.1996
Published
1996-10-00
Pages
5627-35
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178400
Subset
IM
Databases
GENBANK
L12038, U38241
External Links
PubMed source DOI: 10.1128/jb.178.19.5627-5635.1996 PMC Full Text View journal details
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