Single-copy locus proteomics of early- and late-firing DNA replication origins identifies a role of Ask1/DASH complex in replication timing control

• DOI: 10.1016/j.celrep.2023.112045
• Publication Date: 2023-01-26T00:21:29Z
• Authors: Matthias Weiβ; Anna Chanou; Tamas Schauer; Andrey Tvardovskiy; Stefan Meiser; Ann-Christine König; Tobias Schmidt; Elisabeth Kruse; Henning Ummethum; Manuel Trauner; Marcel Werner; Maxime Lalonde; Stefanie M. Hauck; Antonio Scialdone; Stephan Hamperl
• Keywords: Replication timing; Replication; Pre-replication complex; Replication factor C; Origin recognition complex

The chromatin environment at origins of replication is thought to influence DNA initiation in eukaryotic genomes. However, it remains unclear how and which features control the firing early-efficient (EE) or late-inefficient (LI) origins. Here, we use site-specific recombination single-locus isolation purify EE LI Saccharomyces cerevisiae. Using mass spectrometry, define protein composition native regions surrounding start sites. In addition known origin interactors, find microtubule-binding Ask1/DASH complex as an origin-regulating factor. Strikingly, tethering Ask1 individual sites advances timing (RT) targeted chromosomal domain. Targeted degradation globally changes RT a subset origins, can be reproduced by inhibiting microtubule dynamics. Thus, our findings mechanistically connect organization via with cytoskeleton.