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Characterization of Bacillus subtilis ExoA protein: a multifunctional DNA-repair enzyme similar to the Escherichia coli exonuclease III
- Source :
- Bioscience, biotechnology, and biochemistry. 63(9)
- Publication Year :
- 1999
-
Abstract
- To discover the physiological role of the Bacillus subtilis ExoA protein, which is similar in amino acid sequence to Escherichia coli exonuclease III, an exoA::Cm disruption was constructed in the chromosomal DNA of B. subtilis. There was no clear difference in tolerance to hydrogen peroxide and alkylating agents between the disruptant and the wild type strain. An expression plasmid of the ExoA in E. coli was constructed by inserting the exoA gene into the expression vector pKP1500. The purified ExoA was used to clarify enzymatic characterizations using synthetic DNA oligomers as substrates. A DNA oligomer containing a 1', 2'-dideoxyribose residue as an AP site, a DNA-RNA chimera oligomer, and a 3' end 32P-labeled oligomer were synthesized. It has been shown that the ExoA has AP endonuclease, 3'-5' exonuclease, ribonuclease H, and 3'-phosphomonoesterase activities. Thus, it has been confirmed that ExoA is a multifunctional DNA-repair enzyme in B. subtilis that is very similar to E. coli exonuclease III except that ExoA has lower 3'-5' exonuclease activity than that of E. coli exonuclease III.
- Subjects :
- Exonuclease
Exodeoxyribonuclease V
DNA Ligases
Carbon-Oxygen Lyases
Ribonuclease H
Bacillus subtilis
medicine.disease_cause
Applied Microbiology and Biotechnology
Biochemistry
Analytical Chemistry
AP endonuclease
chemistry.chemical_compound
Multienzyme Complexes
medicine
DNA-(Apurinic or Apyrimidinic Site) Lyase
Escherichia coli
AP site
Molecular Biology
DNA Primers
Exonuclease III
Expression vector
biology
Base Sequence
Escherichia coli Proteins
Organic Chemistry
General Medicine
biology.organism_classification
Molecular biology
Deoxyribonuclease IV (Phage T4-Induced)
Recombinant Proteins
Exodeoxyribonucleases
chemistry
Mutation
biology.protein
DNA
Biotechnology
Subjects
Details
- ISSN :
- 09168451
- Volume :
- 63
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Accession number :
- edsair.doi.dedup.....c9e3db46b3e9f541d6c23f31358dbdd3