A methyl-TROSY approach for NMR studies of high-molecular-weight DNA with application to the nucleosome core particle

  • Gili Abramov
    Department of Molecular Genetics, The University of Toronto, Toronto, ON M5S 1A8, Canada;
  • Algirdas Velyvis
    Department of Molecular Genetics, The University of Toronto, Toronto, ON M5S 1A8, Canada;
  • Enrico Rennella
    Department of Molecular Genetics, The University of Toronto, Toronto, ON M5S 1A8, Canada;
  • Leo E. Wong
    Department of Molecular Genetics, The University of Toronto, Toronto, ON M5S 1A8, Canada;
  • Lewis E. Kay
    Department of Molecular Genetics, The University of Toronto, Toronto, ON M5S 1A8, Canada;

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<jats:title>Significance</jats:title> <jats:p>Many of the important functions in the cell are performed by high-molecular-weight complexes that contain both protein and nucleic acid components. Although developments in solution NMR spectroscopy have facilitated the study of the protein portions of these complexes, similar quantitative studies of the nucleic acids have proven challenging. Here we describe an approach for labeling DNA molecules with methyl-group probes at specific sites, followed by the use of optimized NMR methods to obtain high-quality data in studies of these large molecular complexes. Studies of both the DNA and protein components of the nucleosome core particle are presented, including an investigation of how CpG methylation, an important epigenetic mark, affects the structural dynamics of the particle.</jats:p>

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