Extended Data Fig. 8: Chl1 helicase stimulates cohesin loading through interaction with Ctf4.
From: Coordination of cohesin and DNA replication observed with purified proteins
a, Native agarose gel image and b, quantitative comparison of the Order 1 reaction as in Fig. 4b but with omission of Scc2-Scc4, expressed relative to the +Scc2-Scc4 / −Chl1 reaction (denoted by stars, n = 3 independent experiments; means ± s.d.). c, Representative gel image of the experiments in Fig. 4e (n = 3 independent experiments). d, Input fractions of the cohesin-IP experiments of Fig. 4f were analysed by SDS-PAGE and oriole staining (n = 2 independent experiments). e, Order 1 reactions were performed in the absence of the indicated proteins. f, Quantitative comparison of e, expressed relative to the None / −Chl1 reaction (denoted by stars, n = 3 independent experiments; means ± s.d.). g, Order 1 reactions were performed in the presence of the indicated proteins, with the addition of Polδ, Fen1 and ligaseCdc9. h, Quantitative comparison of g, expressed relative to the None / −Chl1 reaction (denoted by stars, n = 3 independent experiments; means ± s.d.). i, Wild-type Chl1, but not the K48R mutant, displays ATP-dependent helicase activity. After incubation with Y-fork structured DNA, the unwound products were detected by native PAGE. The reactions were carried out in 1 mM of ATP and Mg(oAc)2 and 50 mM NaCl. A representative result from two independent experiments is shown. j, Representative gel image of the experiments in Fig. 4g (n = 3 independent experiments). Note that Polδ was omitted from the in vitro replication reactions unless otherwise indicated.
