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Bacterial ribonuclease: mutagenic effect in microbial test-systems

O Ilinskaya1, O B Ivanchenko, N S Karamova

  • 1Microbiology Department, Kazan State University, Tatarstan, Russia.

Mutagenesis
|May 1, 1995
PubMed
Summary

Bacillus intermedius 7P ribonuclease (binase) showed weak mutagenic effects in microbial tests but did not directly damage DNA. Its genotoxicity is recA-dependent, possibly due to nucleotide imbalance from catalytic action on cellular RNAs.

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Area of Science:

  • Microbiology
  • Genotoxicity Testing
  • Enzymology

Background:

  • Bacillus intermedius 7P ribonuclease (binase) is an enzyme with potential biological activity.
  • Investigating the genotoxicity of exogenous enzymes is crucial for understanding their safety and mechanisms of action.

Purpose of the Study:

  • To evaluate the genotoxic potential of purified binase using a battery of microbial assays.
  • To elucidate the mechanism underlying any observed genotoxicity, including the role of enzyme activity and DNA repair pathways.

Main Methods:

  • Utilized four microbial genotoxicity tests: Ames test, AraR-assay, prophage induction, and DNA-repair assays.
  • Assessed mutagenicity with and without metabolic activation using various liver and plant microsomal fractions.
  • Investigated genotoxicity in different bacterial strains deficient in DNA repair mechanisms (uvrA, polA, recA).

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Main Results:

  • Binase exhibited weak mutagenicity at high concentrations, inducing frameshift and base pair substitution mutations.
  • Metabolic activation abolished binase-induced mutagenicity.
  • Binase demonstrated DNA damaging activity in specific mutant strains and induced recA-dependent mutagenesis in Bacillus subtilis and Streptomyces lavendulae.

Conclusions:

  • Binase is not a direct genotoxic agent but can induce recA-dependent mutagenesis.
  • The genotoxic effect is likely mediated by catalytic action on cellular RNAs, leading to nucleotide imbalance and DNA replication alterations.
  • Both catalytic activity and native structure of the enzyme are important for its observed genotoxic effects.