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PMID: 9393705 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

The arginine deiminase pathway in Rhizobium etli: DNA sequence analysis and functional study of the arcABC genes.

Journal of bacteriology ·Vol. 179 ·No. 23 ·1997-12-00 ·Pages 7403-9

D'Hooghe I, Vander Wauven C, Michiels J, Tricot C, de Wilde P, Vanderleyden J, Stalon V

Abstract

Sequence analysis upstream of the Rhizobium etli fixLJ homologous genes revealed the presence of three open reading frames homologous to the arcABC genes of Pseudomonas aeruginosa. The P. aeruginosa arcABC genes code for the enzymes of the arginine deiminase pathway: arginine deiminase, catabolic ornithine carbamoyltransferase (cOTCase), and carbamate kinase. OTCase activities were measured in free-living R. etli cells and in bacteroids isolated from bean nodules. OTCase activity in free-living cells was observed at a different pH optimum than OTCase activity in bacteroids, suggesting the presence of two enzymes with different characteristics and different expression patterns of the corresponding genes. The characteristics of the OTCase isolated from the bacteroids were studied in further detail and were shown to be similar to the properties of the cOTCase of P. aeruginosa. The enzyme has a pH optimum of 6.8 and a molecular mass of approximately 450 kDa, is characterized by a sigmoidal carbamoyl phosphate saturation curve, and exhibits a cooperativity for carbamoyl phosphate. R. etli arcA mutants, with polar effects on arcB and arcC, were constructed by insertion mutagenesis. Bean nodules induced by arcA mutants were still able to fix nitrogen but showed a significantly lower acetylene reduction activity than nodules induced by the wild type. No significant differences in nodule dry weight, plant dry weight, and number of nodules were found between the wild type and the mutants. Determination of the OTCase activity in extracts from bacteroids revealed a strong decrease in activity of this enzyme in the arcA mutant compared to the wild-type strain. Finally, we observed that expression of an R. etli arcA-gusA fusion was strongly induced under anaerobic conditions.

MeSH Terms
Acetylene/metabolism Amino Acid Sequence Fabaceae/microbiology Gene Expression Regulation, Bacterial Genes, Bacterial Hydrolases/biosynthesis,genetics Molecular Sequence Data Ornithine Carbamoyltransferase/genetics Phosphotransferases (Carboxyl Group Acceptor)/genetics Plants, Medicinal Recombinant Fusion Proteins/biosynthesis Rhizobium/enzymology,genetics Sequence Analysis, DNA Sequence Homology, Amino Acid
Chemicals
Recombinant Fusion Proteins Ornithine Carbamoyltransferase Phosphotransferases (Carboxyl Group Acceptor) carbamate kinase Hydrolases arginine deiminase Acetylene
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
D'Hooghe I
F. A. Janssens Laboratory of Genetics, Katholieke Universiteit Leuven, Heverlee, Belgium.
Vander Wauven C
Michiels J
Tricot C
de Wilde P
Vanderleyden J
Stalon V
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.23.7403-7409.1997
Published
1997-12-00
Pages
7403-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179691
Subset
IM
Databases
GENBANK
AF025543
External Links
PubMed source DOI: 10.1128/jb.179.23.7403-7409.1997 PMC Full Text View journal details
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