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PMID: 2407723 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Analysis of the Escherichia coli gene encoding L-asparaginase II, ansB, and its regulation by cyclic AMP receptor and FNR proteins.

Journal of bacteriology ·Vol. 172 ·No. 3 ·1990-03-00 ·Pages 1491-8

Jennings MP, Beacham IR

Abstract

Escherichia coli contains two L-asparaginase isozymes: L-asparaginase I, a low-affinity enzyme located in the cytoplasm, and L-asparaginase II, a high-affinity secreted enzyme. A molecular genetic analysis of the gene (ansA) encoding the former enzyme has previously been reported. We now present a molecular study of the gene, ansB, encoding L-asparaginase II. This gene was isolated by using oligonucleotide probes, whose sequences were based on the previously determined amino acid sequence. The nucleotide sequence of ansB, including 5'- and 3'-untranslated regions, was determined. The amino acid sequence of L-asparaginase II, deduced from this nucleotide sequence, contains differences at 11 positions when compared with the previously determined amino acid sequence. The deduced amino acid sequence also reveals a typical secretory signal peptide of 22 residues. A single region of sequence similarity is observed when ansA and ansB are compared. The transcriptional start site in ansB was determined, allowing the identification of the promoter region. The regulation of ansB was studied by using ansB'-'lacZ fusions, together with a deletion analysis of the 5' region upstream of the promoter. Regulation by cyclic AMP receptor protein and anaerobiosis (FNR protein) was confirmed, and the presence of nucleotide sequence motifs, with homology to cyclic AMP receptor protein and FNR protein-binding sites, investigated.

MeSH Terms
Amino Acid Sequence Asparaginase/genetics Bacterial Proteins/metabolism Base Sequence Blotting, Southern Cloning, Molecular DNA, Recombinant/metabolism Escherichia coli/enzymology,genetics Escherichia coli Proteins Gene Expression Regulation, Bacterial Gene Expression Regulation, Enzymologic Gene Library Iron-Sulfur Proteins Isoenzymes/genetics Molecular Sequence Data Nucleic Acid Hybridization Plasmids Receptors, Cyclic AMP/metabolism Restriction Mapping Sequence Homology, Nucleic Acid Transcription Factors/metabolism Transcription, Genetic
Chemicals
Bacterial Proteins DNA, Recombinant Escherichia coli Proteins FNR protein, E coli Iron-Sulfur Proteins Isoenzymes Receptors, Cyclic AMP Transcription Factors Asparaginase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Jennings M P
Division of Science and Technology, Griffith University, Brisbane, Queensland, Australia.
Beacham I R
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.3.1491-1498.1990
Published
1990-03-00
Pages
1491-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208625
Subset
IM
Databases
GENBANK
M34277
External Links
PubMed source DOI: 10.1128/jb.172.3.1491-1498.1990 PMC Full Text View journal details
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