Journal Articles

The Folding of the Specific DNA Recognition Subdomain of the Sleeping Beauty Transposase Is Temperature-Dependent and Is Required for Its Binding to the Transposon DNA

November 06, 2014

The Folding of the Specific DNA Recognition Subdomain of the Sleeping Beauty Transposase Is Temperature-Dependent and Is Required for Its Binding to the Transposon DNAPLoS One. 2014 Nov 6, Leighton GO1, Konnova TA2, Idiyatullin B3, Hurr SH1, Zuev YF3, Nesmelova IV4.

The NMR experiments for this study were carried out at the David H. Murdock Research Institute, NC Reseasrch Campus in Kannapolis, NC.

  • 1Department of Physics and Optical Science, University of North Carolina, Charlotte, North Carolina, United States of America.
  • 2Department of Physics and Optical Science, University of North Carolina, Charlotte, North Carolina, United States of America; Kazan Institute of Biochemistry and Biophysics, Kazan, Russian Federation.
  • 3Kazan Institute of Biochemistry and Biophysics, Kazan, Russian Federation.
  • 4Department of Physics and Optical Science, University of North Carolina, Charlotte, North Carolina, United States of America; Center for Biomedical Engineering and Science, University of North Carolina, Charlotte, North Carolina, United States of America.
  • FW: Scholar Alert – [ Kannapolis ] – Outlook Web Access Light

 

Abstract

The reaction of DNA transposition begins when the transposase enzyme binds to the transposon DNA. Sleeping Beauty is a member of the mariner family of DNA transposons. Although it is an important tool in application genetic ions and has been adapted for human gene therapy, its molecular mechanism remains obscure. Here, we show that only the folded conformation of the specific DNA recognition subdomain of the Sleeping Beautytransposase, the PAI subdomain, binds to the transposon DNA. Furthermore, we show that the PAI subdomain is well folded at low temperatures, but the presence of unfolded conformation gradually increases at temperatures above 15°C, suggesting that the choice of temperature may be important for the optimal transposase activity. Overall, the results provide a molecular-level insight into the DNA recognition by the Sleeping Beautytransposase.

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