Tracking replication enzymology in vivo by genome-wide mapping of ribonucleotide incorporation View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2015-03

AUTHORS

Anders R Clausen, Scott A Lujan, Adam B Burkholder, Clinton D Orebaugh, Jessica S Williams, Maryam F Clausen, Ewa P Malc, Piotr A Mieczkowski, David C Fargo, Duncan J Smith, Thomas A Kunkel

ABSTRACT

Ribonucleotides are frequently incorporated into DNA during replication in eukaryotes. Here we map genome-wide distribution of these ribonucleotides as markers of replication enzymology in budding yeast, using a new 5' DNA end-mapping method, hydrolytic end sequencing (HydEn-seq). HydEn-seq of DNA from ribonucleotide excision repair-deficient strains reveals replicase- and strand-specific patterns of ribonucleotides in the nuclear genome. These patterns support the roles of DNA polymerases α and δ in lagging-strand replication and of DNA polymerase ɛ in leading-strand replication. They identify replication origins, termination zones and variations in ribonucleotide incorporation frequency across the genome that exceed three orders of magnitude. HydEn-seq also reveals strand-specific 5' DNA ends at mitochondrial replication origins, thus suggesting unidirectional replication of a circular genome. Given the conservation of enzymes that incorporate and process ribonucleotides in DNA, HydEn-seq can be used to track replication enzymology in other organisms. More... »

PAGES

185-191

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nsmb.2957

DOI

http://dx.doi.org/10.1038/nsmb.2957

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1028327172

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25622295


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