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.
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 genetic applications 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 Beauty transposase, 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 Beauty transposase.
Department of Physics and Optical Science, University of North Carolina, Charlotte, North Carolina, United States of America.
Gage O
GO
Leighton
Department 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.
Tatiana A
TA
Konnova
Kazan Institute of Biochemistry and Biophysics, Kazan, Russian Federation.
Bulat
B
Idiyatullin
Department of Physics and Optical Science, University of North Carolina, Charlotte, North Carolina, United States of America.
Sophia H
SH
Hurr
Kazan Institute of Biochemistry and Biophysics, Kazan, Russian Federation.
Yuriy F
YF
Zuev
Department 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.
Irina V
IV
Nesmelova
D004251:DNA Transposable Elements / Q000502:physiology; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013696:Temperature; D019895:Transposases / Q000737:chemistry* / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry
D004251:DNA Transposable Elements; D014443:Tyrosine; D019895:Transposases; C494471:sleeping beauty transposase, human