Print Email Facebook Twitter A chromatinized origin reduces the mobility of ORC and MCM through interactions and spatial constraint Title A chromatinized origin reduces the mobility of ORC and MCM through interactions and spatial constraint Author Sanchez González, H. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Liu, Z. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) van Veen, E.N.W. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) van Laar, T. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Diffley, John F.X. (Francis Crick Institute) Dekker, N.H. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Date 2023 Abstract Chromatin replication involves the assembly and activity of the replisome within the nucleosomal landscape. At the core of the replisome is the Mcm2-7 complex (MCM), which is loaded onto DNA after binding to the Origin Recognition Complex (ORC). In yeast, ORC is a dynamic protein that diffuses rapidly along DNA, unless halted by origin recognition sequences. However, less is known about the dynamics of ORC proteins in the presence of nucleosomes and attendant consequences for MCM loading. To address this, we harnessed an in vitro single-molecule approach to interrogate a chromatinized origin of replication. We find that ORC binds the origin of replication with similar efficiency independently of whether the origin is chromatinized, despite ORC mobility being reduced by the presence of nucleosomes. Recruitment of MCM also proceeds efficiently on a chromatinized origin, but subsequent movement of MCM away from the origin is severely constrained. These findings suggest that chromatinized origins in yeast are essential for the local retention of MCM, which may facilitate subsequent assembly of the replisome. To reference this document use: http://resolver.tudelft.nl/uuid:6328df2c-be3e-4db5-b373-8d552f786fee DOI https://doi.org/10.1038/s41467-023-42524-8 ISSN 2041-1723 Source Nature Communications, 14 (1) Part of collection Institutional Repository Document type journal article Rights © 2023 H. Sanchez González, Z. Liu, E.N.W. van Veen, T. van Laar, John F.X. Diffley, N.H. Dekker Files PDF s41467_023_42524_8.pdf 1.77 MB Close viewer /islandora/object/uuid:6328df2c-be3e-4db5-b373-8d552f786fee/datastream/OBJ/view