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PMID: 1569015 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Nucleotide sequence and transcriptional analysis of activator-regulator proteins for beta-lactamase in Streptomyces cacaoi.

Journal of bacteriology ·Vol. 174 ·No. 9 ·1992-05-00 ·Pages 2834-42

Urabe H, Ogawara H

Abstract

The nucleotide sequence of the 2.7-kb DNA fragment upstream of the structural gene of beta-lactamase in Streptomyces cacaoi was determined. Computer-aided "FRAME" analysis revealed four possible open reading frames (ORFs), three in one direction and one in the opposite direction. One of them (ORF1, BlaA) encoded an activator-regulator protein whose deduced amino acid sequence was similar to that of other activator-regulator proteins in bacteria. Insertion of an 8-bp BamHI linker into the BlaA region decreased the beta-lactamase activity sharply, from 50 U to 1 U/ml. This protein (BlaA) was found to bind to the nucleotide sequence between the bla (beta-lactamase structural gene) and blaA genes. Another ORF (ORF2, BlaB) in the same orientation had a couple of amino acid sequences similar to that of pBR322 beta-lactamase. However, insertion of the 8-bp BamHI linker indicated that this ORF was functional as an activator-regulator but not as a beta-lactamase. Therefore, there were two activator-regulator proteins in the upstream region of the structural gene of the beta-lactamase. Nuclease S1 mapping predicted that transcription for the activator proteins commenced at the translational initiation codon or within a few nucleotides from the translational start site. Transcription was in the opposite direction to that of the beta-lactamase structural gene.

MeSH Terms
Amino Acid Sequence Bacterial Proteins Base Sequence Chromosome Mapping DNA Mutational Analysis DNA-Binding Proteins/genetics Genes, Bacterial/genetics Molecular Sequence Data Protein Biosynthesis Reading Frames Regulatory Sequences, Nucleic Acid/genetics Sequence Homology, Nucleic Acid Streptomyces/genetics Transcription, Genetic beta-Lactamases/biosynthesis,genetics
Chemicals
Bacterial Proteins DNA-Binding Proteins BlaA protein, Bacteria beta-Lactamases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Urabe H
Department of Biochemistry, Meiji College of Pharmacy, Tokyo, Japan.
Ogawara H
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.9.2834-2842.1992
Published
1992-05-00
Pages
2834-42
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205934
Subset
IM
Databases
GENBANK
D00937, M79367, M79368, M79369, M79370, M79371, M79372, M81469, M89479, M89480
External Links
PubMed source DOI: 10.1128/jb.174.9.2834-2842.1992 PMC Full Text View journal details
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