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Stabilization of the primary sigma factor of Staphylococcus aureus by core RNA polymerase

  • Mondal, Rajkrishna (Department of Biochemistry, Bose Institute) ;
  • Ganguly, Tridib (School of Life Sciences, IISER) ;
  • Chanda, Palas K. (Department of Biochemistry, Bose Institute) ;
  • Bandhu, Amitava (Department of Biochemistry, Bose Institute) ;
  • Jana, Biswanath (Department of Biochemistry, Bose Institute) ;
  • Sau, Keya (School of Biotechnology and Life Sciences, Haldia Institute of Technology) ;
  • Lee, Chia-Y. (Department of Microbiology and Immunology, University of Arkansas for Medical Sciences) ;
  • Sau, Subrata (Department of Biochemistry, Bose Institute)
  • 발행 : 2010.03.31

초록

The primary sigma factor ($\sigma^{A}$) of Staphylococcus aureus, a potential drug target, was little investigated at the structural level. Using an N-terminal histidine-tagged $\sigma^{A}$ (His-$\sigma^{A}$), here we have demonstrated that it exits as a monomer in solution, possesses multiple domains, harbors primarily $\alpha$-helix and efficiently binds to a S. aureus promoter DNA in the presence of core RNA polymerase. While both N- and C-terminal ends of His-$\sigma^{A}$ are flexible in nature, two Trp residues in its DNA binding region are buried. Upon increasing the incubation temperature from 25$^{\circ}$ to 40$^{\circ}C$, $\sim$60% of the input His-$\sigma^{A}$ was cleaved by thermolysin. Aggregation of His-$\sigma^{A}$ was also initiated rapidly at 45$^{\circ}C$. From the equilibrium unfolding experiment, the Gibbs free energy of stabilization of His-$\sigma^{A}$ was estimated to be +0.70 kcal $mol^{-1}$. The data together suggest that primary sigma factor of S. aureus is an unstable protein. Core RNA polymerase however stabilized $\sigma^{A}$ appreciably.

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피인용 문헌

  1. Insights into the regulation of small RNA expression: SarA represses the expression of two sRNAs inStaphylococcus aureus 2016, https://doi.org/10.1093/nar/gkw777
  2. AFM study of Escherichia coli RNA polymerase σ70 subunit aggregation vol.8, pp.1, 2012, https://doi.org/10.1016/j.nano.2011.05.014