31P NMR investigation of backbone dynamics in DNA binding sites.
The backbone conformation of DNA plays an important role in the indirect readout mechanisms for protein--DNA recognition events. Thus, investigating the backbone dynamics of each step in DNA binding sequences provides useful information necessary for the characterization of these interactions. Here, we use 31P dynamic NMR to characterize the backbone conformation and dynamics in the Dickerson dodecamer, a sequence containing the EcoRI binding site, and confirm solid-state 2H NMR results showing that the C3pG4 and C9pG10 steps experience unique dynamics and that these dynamics are quenched upon cytosine methylation. In addition, we show that cytosine methylation affects the conformation and dynamics of neighboring nucleotide steps, but this effect is localized to only near neighbors and base-pairing partners. Last, we have been able to characterize the percent BII in each backbone step and illustrate that the C3pG4 and C9pG10 favor the noncanonical BII conformation, even at low temperatures. Our results demonstrate that 31P dynamic NMR provides a robust and efficient method for characterizing the backbone dynamics in DNA. This allows simple, rapid determination of sequence-dependent dynamical information, providing a useful method for studying trends in protein-DNA recognition events.
Tian
Ye
Y
Department of Chemistry, University of California, Riverside, California 92521, USA.
Kayatta
Michael
M
Shultis
Katharine
K
Gonzalez
Alejandro
A
Mueller
Leonard J
LJ
Hatcher
Mary E
ME
D000465:Algorithms; D001665:Binding Sites; D001703:Biophysics / Q000379:methods; D004247:DNA / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008962:Models, Theoretical; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry; D010761:Phosphorus Radioisotopes / Q000627:therapeutic use*; D011506:Proteins / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D011506:Proteins; D004247:DNA