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Weiß, M. ; Chanou, A. ; Schauer, T. ; Tvardovskiy, A. ; Meiser, S. ; König, A.-C. ; Schmidt, T. ; Kruse, E. ; Ummethum, H. ; Trauner, M. ; Werner, M. ; Lalonde, M. ; Hauck, S.M. ; Scialdone, A. ; Hamperl, S.

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

Cell Rep. 42:112045 (2023)

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Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.

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

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Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Ask1 Protein ; Cp: Molecular Biology ; Dash Complex ; Dna Replication Origin ; Affinity Purification ; Locus-specific Chromatin Isolation ; Microtubule Cytoskeleton ; Origin Chromatin Structure ; Replication Efficiency ; Replication Timing ; Site-specific Recombination; Kinetochore-microtubule Interface; Yeast; Chromatin; Binding; Recognition; Activation; Initiation; Sequence; Dynamics; Time

ISSN (print) / ISBN 2211-1247

e-ISSN 2211-1247

Journal Cell Reports

Quellenangaben Volume: 42, Issue: 2, Article Number: 112045

Publisher Cell Press

Publishing Place 50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Epigenetics and Stem Cells (IES)
Institute of Functional Epigenetics (IFE)
CF Metabolomics & Proteomics (CF-MPC)
Institute of Computational Biology (ICB)

Grants
Helmholtz Gesellschaft
European Research Council (ERC starting grant)
DFG