In vitro Assays for Eukaryotic Leading/Lagging Strand DNA Replication.

Content Provider	Europe PMC
Author	Schauer, Grant Finkelstein, Jeff O’Donnell, Mike
Copyright Year	2017
Abstract	The eukaryotic replisome is a multiprotein complex that duplicates DNA. The replisome is sculpted to couple continuous leading strand synthesis with discontinuous lagging strand synthesis, primarily carried out by DNA polymerases ε and δ, respectively, along with helicases, polymerase α-primase, DNA sliding clamps, clamp loaders and many other proteins. We have previously established the mechanisms by which the polymerases ε and δ are targeted to their ‘correct’ strands, as well as quality control mechanisms that evict polymerases when they associate with an ‘incorrect’ strand. Here, we provide a practical guide to differentially assay leading and lagging strand replication in vitro using pure proteins.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC5659624&blobtype=pdf
Volume Number	7
PubMed Central reference number	PMC5659624
Issue Number	18
PubMed reference number	29082290
Journal	Bio-protocol [Bio Protoc]
e-ISSN	23318325
DOI	10.21769/bioprotoc.2548
Language	English
Publisher	Bio-Protocol
Publisher Date	2017-09-01
Publisher Place	1075 Lorne Way, Sunnyvale, CA 94087, USA
Access Restriction	Open
Rights License	Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC.
Subject Keyword	Eukaryotic DNA replication Replisome assay CMG helicase DNA polymerase RFC clamp loader PCNA sliding clamp Leading strand Lagging strand
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Plant Science Biochemistry, Genetics and Molecular Biology