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

ABSTRACT: The development of methyl-transverse relaxation-optimized spectroscopy (methyl-TROSY)-based NMR methods, in concert with robust strategies for incorporation of methyl-group probes of structure and dynamics into the protein of interest, has facilitated quantitative studies of high-molecular-weight protein complexes. Here we develop a one-pot in vitro reaction for producing NMR quantities of methyl-labeled DNA at the C5 and N6 positions of cytosine (5mC) and adenine (6mA) nucleobases, respectively, enabling the study of high-molecular-weight DNA molecules using TROSY approaches originally developed for protein applications. Our biosynthetic strategy exploits the large number of naturally available methyltransferases to specifically methylate DNA at a desired number of sites that serve as pro

SUBMITTER: Abramov G 

PROVIDER: S-EPMC7293644 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature