Expression and mutational analysis of DinB-like protein DR0053 in Deinococcus radiodurans

PLoS One. 2015 Feb 23;10(2):e0118275. doi: 10.1371/journal.pone.0118275. eCollection 2015.

Abstract

In order to understand the mechanism governing radiation resistance in Deinococcus radiodurans, current efforts are aimed at identifying potential candidates from a large repertoire of unique Deinococcal genes and protein families. DR0053 belongs to the DinB/YfiT protein family, which is an over-represented protein family in D. radiodurans. We observed that dr0053 transcript levels were highly induced in response to gamma radiation (γ-radiation) and mitomycin C (MMC) exposure depending on PprI, RecA and the DrtR/S two-component signal transduction system. Protein profiles demonstrated that DR0053 is a highly induced protein in cultures exposed to 10 kGy γ-radiation. We were able to determine the transcriptional start site of dr0053, which was induced upon irradiation, and to assign the 133-bp promoter region of dr0053 as essential for radiation responsiveness through primer extension and promoter deletion analyses. A dr0053 mutant strain displayed sensitivity to γ-radiation and MMC exposure, but not hydrogen peroxide, suggesting that DR0053 helps cells recover from DNA damage. Bioinformatic analyses revealed that DR0053 is similar to the Bacillus subtilis protein YjoA, which is a substrate of bacterial protein-tyrosine kinases. Taken together, the DNA damage-inducible (din) gene dr0053 may be regulated at the transcriptional and post-translational levels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • DNA Damage / drug effects
  • DNA Damage / genetics
  • DNA Mutational Analysis / methods
  • Deinococcus / drug effects
  • Deinococcus / genetics*
  • Gamma Rays
  • Gene Expression Regulation, Bacterial / drug effects
  • Gene Expression Regulation, Bacterial / genetics*
  • Hydrogen Peroxide / pharmacology
  • Mitomycin / pharmacology
  • Mutation / drug effects
  • Mutation / genetics*
  • Promoter Regions, Genetic / genetics
  • Protein-Tyrosine Kinases / genetics
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / genetics

Substances

  • Bacterial Proteins
  • Mitomycin
  • Hydrogen Peroxide
  • Protein-Tyrosine Kinases

Grants and funding

This research was supported by the Nuclear Research and Development Program of the Ministry of Science, ICT, and Future Planning (MSIP), Republic of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.