Site-Directed Mutagenesis of a Catabolite Repression Operator Sequence in Bacillus subtilis

Catabolite repression of the Bacillus subtilis α-amylase gene (amyE) involves an operator sequence located just downstream of the promoter (amyR), overlapping the transcription start site. Oligonucleotide site-directed mutagenesis of this sequence identified bases required for catabolite repression....

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Bibliographic Details
Published inProceedings of the National Academy of Sciences - PNAS Vol. 87; no. 16; pp. 6238 - 6242
Main Authors Weickert, Michael J., Chambliss, Glenn H.
Format Journal Article
LanguageEnglish
Published Washington, DC National Academy of Sciences of the United States of America 01.08.1990
National Acad Sciences
Subjects
alpha-Amylases - biosynthesis
alpha-Amylases - genetics
Bacillus
Bacillus subtilis - genetics
Base Sequence
Biological and medical sciences
Cloning, Molecular
DNA
DNA, Bacterial - genetics
Enzyme Repression
Enzymes
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetic mutation
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Mutagenesis. Repair
Mutation
Oligonucleotide Probes
Oligonucleotides
Operator regions
Operon
Plasmids
Promoter regions
Promoter Regions, Genetic
Repression
Restriction Mapping
Sequence Homology, Nucleic Acid
Transcription initiation site
Regulation(control)
Bacillales
Gene
Nucleotide sequence
Transcription
Bacillus subtilis
Enzyme
Site directed mutagenesis
Bacteria
α-Amylase
Bacillaceae
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Summary:Catabolite repression of the Bacillus subtilis α-amylase gene (amyE) involves an operator sequence located just downstream of the promoter (amyR), overlapping the transcription start site. Oligonucleotide site-directed mutagenesis of this sequence identified bases required for catabolite repression. Two mutations increased both the 2-fold symmetry of the operator and the repression ratio. Although many mutations reduced the repression ratio 3- to 11-fold, some also caused a 2-fold or greater increase in amylase production. Others caused hyperproduction without affecting catabolite repression. Homologous sequences in other catabolite-repressed B. subtilis promoters suggest a common regulatory site may be involved in catabolite repression.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.87.16.6238