Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation

• DOI: 10.7554/elife.04988
• Publication Date: 2015-04-14T11:34:31Z
• Authors: Roxana E Georgescu; Grant D Schauer; Nina Y Yao; Lance D Langston; Olga Yurieva; Dan Zhang; Jeff Finkelstein; Mike E O'Donnell
• Keywords: Replisome; Okazaki fragments

We have reconstituted a eukaryotic leading/lagging strand replisome comprising 31 distinct polypeptides. This study identifies process unprecedented in bacterial replisomes. While bacteria and phage simply recruit polymerases to the fork, we find that suppression mechanisms are used position on their respective strands. Hence, Pol ε is active with CMG leading strand, but it unable function lagging even when δ not present. Conversely, δ-PCNA only enzyme capable of extending Okazaki fragments presence Pols α. shown earlier suppressed (<xref ref-type="bibr" rid="bib12">Georgescu et al., 2014</xref>). propose CMG, 11-subunit helicase, responsible for one or both these spatially control polymerase occupancy at fork.