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PMID: 3131308 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Anabolic ornithine carbamoyltransferase of Pseudomonas aeruginosa: nucleotide sequence and transcriptional control of the argF structural gene.

Journal of bacteriology ·Vol. 170 ·No. 6 ·1988-06-00 ·Pages 2725-34

Itoh Y, Soldati L, Stalon V, Falmagne P, Terawaki Y, Leisinger T, Haas D

Abstract

In Pseudomonas aeruginosa PAO the anabolic ornithine carbamoyltransferase (OTCase, EC 2.1.3.3) is the product of the argF gene and the only arginine biosynthetic enzyme whose synthesis is repressible by arginine. We have determined the complete nucleotide sequence of the argF gene including its promoter-control region. The deduced amino acid sequence of the anabolic OTCase consists of 305 residues (Mr 33,924), and this was confirmed by the N-terminal amino acid sequence, the total amino acid composition, and the subunit Mr of the purified enzyme. The native anabolic OTCase (Mr 110,000 to 125,000) was found to be a trimer by cross-linking experiments. P. aeruginosa also has a catabolic OTCase (the arcB gene product), which catalyzes the reverse reaction of the anabolic conversion. At the nucleotide sequence level, the P. aeruginosa argF gene had 52.4% identity with the arcB gene. The Escherichia coli argF and argI genes, which code for anabolic OTCase isoenzymes, had 47.3 and 44.9% identity, respectively, with the P. aeruginosa argF sequence. This suggests that these four genes have evolved from a common ancestral gene. The arcB gene appears to be more closely related to the E. coli argF gene than to the P. aeruginosa argF gene. Two transcripts (mRNA-1, mRNA-2) of the P. aeruginosa argF gene were identified by S1 mapping. The transcription initiation site for mRNA-1 was preceded by sequences having partial homology with the E. coli -35 and -10 consensus promoter sequences. No sequence similar to consensus promoters of enteric bacteria was found upstream of the 5' end of mRNA-2. E. coli carrying a P. aeruginosa argF+ recombinant plasmid produced mRNA-1 with low efficiency but no (or very little) mRNA-2. Arginine repressed argF transcription in P. aeruginosa. In the argF promoter region no sequence homologous to the "arg box" (arginine operator module) of E. coli was found. The mechanism of arginine repression in P. aeruginosa thus appears to be different from that in E. coli.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Gene Expression Regulation Genes Molecular Sequence Data Ornithine Carbamoyltransferase/genetics Promoter Regions, Genetic Pseudomonas aeruginosa/enzymology,genetics Transcription, Genetic
Chemicals
Ornithine Carbamoyltransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Itoh Y
Department of Bacteriology, Shinshu University School of Medicine, Matsumoto, Japan.
Soldati L
Stalon V
Falmagne P
Terawaki Y
Leisinger T
Haas D
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.170.6.2725-2734.1988
Published
1988-06-00
Pages
2725-34
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC211195
Subset
IM
Databases
GENBANK
M19939
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1128/jb.170.6.2725-2734.1988 PMC Full Text View journal details
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