Extended Data Fig. 7: Replication-specific stimulation of cohesin loading by Chl1.

a, Schematic of the Order 1 reaction using the chromatinized circular DNA templates. b, Native agarose gel analysis of the experiments performed as depicted in a. A representative result from two independent experiments is shown. c, Schematic of the Order 1 reaction but including Pif1 and post-replicative restriction digestion. Pfi1 was added to the reaction 10 min after replication initiation. See Methods for details. d, Native agarose gel analysis and e, quantitative comparison of the experiments performed as depicted in c, expressed relative to the −Pif1 / −Chl1 reaction (denoted by stars). Means ± s.d. from three independent experiments are shown. f, Chl1 was added in the “cohesin first” reaction as denoted in the schematic. g, Native agarose gel analysis and h, quantitative comparison of the experiments performed as depicted in f, expressed relative to the −Pif1 / −Chl1 reaction (denoted by stars). Means ± s.d. from three independent experiments are shown. i, Cohesin loading reactions were carried out with the indicated cohesin and Scc2-Scc4 concentrations, and then replication was initiated in the absence or presence of Chl1. j, Native agarose gel analysis and k, quantitative comparison of the experiments performed as depicted in i. Means from two independent experiments are shown. l, Chl1 was added in the “cohesin first” reaction using the chromatinized template as denoted in the schematic. m, Native agarose gel analysis and n, quantitative comparison of the experiments performed as depicted in l. Means of two independent experiments are shown. Note that Polδ was omitted from the in vitro replication reactions using naked DNA to avoid the occurrence of strand displacement synthesis by Polδ, which is pronounced by Pif1³², whereas Pol δ was included in chromatin replication because nucleosomes limit the strand displacement synthesis^37,38.

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