High resolution mapping of modified DNA nucleobases using excision repair enzymes.

Abstract:

:The incorporation and creation of modified nucleobases in DNA have profound effects on genome function. We describe methods for mapping positions and local content of modified DNA nucleobases in genomic DNA. We combined in vitro nucleobase excision with massively parallel DNA sequencing (Excision-seq) to determine the locations of modified nucleobases in genomic DNA. We applied the Excision-seq method to map uracil in E. coli and budding yeast and discovered significant variation in uracil content, wherein uracil is excluded from the earliest and latest replicating regions of the genome, possibly driven by changes in nucleotide pool composition. We also used Excision-seq to identify sites of pyrimidine dimer formation induced by UV light exposure, where the method could distinguish between sites of cyclobutane and 6-4 photoproduct formation. These UV mapping data enabled analysis of local sequence bias around pyrimidine dimers and suggested a preference for an adenosine downstream from 6-4 photoproducts. The Excision-seq method is broadly applicable for high precision, genome-wide mapping of modified nucleobases with cognate repair enzymes.

journal_name

Genome Res

journal_title

Genome research

authors

Bryan DS,Ransom M,Adane B,York K,Hesselberth JR

doi

10.1101/gr.174052.114

subject

Has Abstract

pub_date

2014-09-01 00:00:00

pages

1534-42

issue

9

eissn

1088-9051

issn

1549-5469

pii

gr.174052.114

journal_volume

24

pub_type

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