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Extremophiles

, Volume 20, Issue 2, pp 195–205 | Cite as

DNA Gyrase of Deinococcus radiodurans is characterized as Type II bacterial topoisomerase and its activity is differentially regulated by PprA in vitro

  • Swathi Kota
  • Yogendra S. Rajpurohit
  • Vijaya K. Charaka
  • Katsuya Satoh
  • Issay Narumi
  • Hari S. MisraEmail author
Original Paper

Abstract

The multipartite genome of Deinococcus radiodurans forms toroidal structure. It encodes topoisomerase IB and both the subunits of DNA gyrase (DrGyr) while lacks other bacterial topoisomerases. Recently, PprA a pleiotropic protein involved in radiation resistance in D. radiodurans has been suggested for having roles in cell division and genome maintenance. In vivo interaction of PprA with topoisomerases has also been shown. DrGyr constituted from recombinant gyrase A and gyrase B subunits showed decatenation, relaxation and supercoiling activities. Wild type PprA stimulated DNA relaxation activity while inhibited supercoiling activity of DrGyr. Lysine133 to glutamic acid (K133E) and tryptophane183 to arginine (W183R) replacements resulted loss of DNA binding activity in PprA and that showed very little effect on DrGyr activities in vitro. Interestingly, wild type PprA and its K133E derivative continued interacting with GyrA in vivo while W183R, which formed relatively short oligomers did not interact with GyrA. The size of nucleoid in PprA mutant (1.9564 ± 0.324 µm) was significantly bigger than the wild type (1.6437 ± 0.345 µm). Thus, we showed that DrGyr confers all three activities of bacterial type IIA family DNA topoisomerases, which are differentially regulated by PprA, highlighting the significant role of PprA in DrGyr activity regulation and genome maintenance in D. radiodurans.

Keywords

Deinococcus DNA gyrase Genome maintenance PprA Radioresistance 

Notes

Acknowledgments

We thank Dr. S. Chattopadhyay for his comments and supports and Drs Kruti Mehta-Modi, Anubrata Das and Chitra S Misra for their editorial and technical comments in this manuscript.

Supplementary material

792_2016_814_MOESM1_ESM.pdf (563 kb)
Supplementary material 1 (PDF 562 kb)

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Copyright information

© Springer Japan 2016

Authors and Affiliations

  • Swathi Kota
    • 1
  • Yogendra S. Rajpurohit
    • 1
  • Vijaya K. Charaka
    • 1
    • 4
  • Katsuya Satoh
    • 2
  • Issay Narumi
    • 3
  • Hari S. Misra
    • 1
    Email author
  1. 1.Molecular Biology DivisionBhabha Atomic Research CentreMumbaiIndia
  2. 2.Ion Beam Mutagenesis Research Group, Quantum Beam Science CenterJapan Atomic Energy AgencyTakasakiJapan
  3. 3.Radiation Microbiology Laboratory, Department of Life SciencesToyo UniversityItakuraJapan
  4. 4.Department of Radiation Oncology, Houston Methodist Research InstituteHouston Methodist HospitalHoustonUSA

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