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Citations

Citations

Thymine DNA glycosylase combines sliding, hopping, and nucleosome interactions to efficiently search for 5-formylcytosine

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Brittani L Schnable, Matthew A Schaich, Vera Roginskaya, Liam P Leary, Tyler M Weaver, Bret D Freudenthal, Alexander C Drohat, Bennett Van Houten
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Gene Synthesis ...Wild type TDG-HaloTag and TDG-HaloTag variants were expressed from plasmids constructed utilizing Gene Universal Inc with the pHTN HaloTag CMV-neo vector (Promega). (... Get A Quote

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...Wild type TDG-HaloTag and TDG-HaloTag variants were expressed from plasmids constructed utilizing Gene Universal Inc with the pHTN HaloTag CMV-neo vector (Promega). (...

Abstract

Base excision repair is the main pathway involved in active DNA demethylation. 5-formylcytosine and 5-carboxylcytosine, two oxidized moieties of methylated cytosine, are recognized and removed by thymine DNA glycosylase (TDG) to generate an abasic site. Using single molecule fluorescence experiments, we study TDG in the presence and absence of 5-formylcytosine. TDG exhibits multiple modes of linear diffusion, including hopping and sliding, in search of base modifications. TDG active site variants and truncated N-terminus, reveals these variants alter base modification search and recognition mechanism of TDG. On DNA containing an undamaged nucleosome, TDG is found to either bypass, colocalize with, or encounter but not bypass the nucleosome. Truncating the N-terminus reduces the number of interactions with the nucleosome. Our findings provide mechanistic insights into how TDG searches for modified DNA bases in chromatin.

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