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DNA replication stress is a prominent endogenous source of genome instability that not only leads to ageing, but also neuropathology and cancer development in humans. Specifically, the issues of how vertebrate cells select and activate origins of replication are of importance as, for example, insufficient origin firing leads to genomic instability and mutations in replication initiation factors lead to the rare human disease Meier-Gorlin syndrome. The mechanism of origin activation has been well characterised and reconstituted in yeast, however, an equal understanding of this process in higher eukaryotes is lacking. The firing of replication origins is driven by S-phase kinases (CDKs and DDK) and results in the activation of the replicative helicase and generation of two bi-directional replication forks. Our data, generated from cell-free Xenopus laevis egg extracts, show that DONSON is required for assembly of the active replicative helicase (CMG complex) at origins during replication initiation. DONSON has previously been shown to be essential during DNA replication, both in human cells and in Drosophila, but the mechanism of DONSON\u2019s action was unknown. Here we show that DONSON\u2019s presence is essential for replication initiation as it is required for Cdc45 and GINS association with Mcm2\u20137 complexes and helicase activation. To fulfil this role, DONSON interacts with the initiation factor, TopBP1, in a CDK-dependent manner. Following its initiation role, DONSON also forms a part of the replisome during the elongation stage of DNA replication. Mutations in DONSON have recently been shown to lead to the Meier-Gorlin syndrome; this novel replication initiation role of DONSON therefore provides the explanation for the phenotypes caused by DONSON mutations in patients.<\/jats:p>","DOI":"10.1093\/nar\/gkad694","type":"journal-article","created":{"date-parts":[[2023,8,9]],"date-time":"2023-08-09T16:45:35Z","timestamp":1691599535000},"page":"9748-9763","source":"Crossref","is-referenced-by-count":45,"title":["DONSON facilitates Cdc45 and GINS chromatin association and is essential for DNA replication initiation"],"prefix":"10.1093","volume":"51","author":[{"given":"Georgia","family":"Kingsley","sequence":"first","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aggeliki","family":"Skagia","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paolo","family":"Passaretti","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cyntia","family":"Fernandez-Cuesta","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicja","family":"Reynolds-Winczura","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kinga","family":"Koscielniak","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4648-0332","authenticated-orcid":false,"given":"Agnieszka","family":"Gambus","sequence":"additional","affiliation":[{"name":"Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,8,28]]},"reference":[{"key":"2023101301523421400_B1","doi-asserted-by":"crossref","first-page":"1194","DOI":"10.1016\/j.cell.2013.05.039","article-title":"The hallmarks of aging","volume":"153","author":"Lopez-Otin","year":"2013","journal-title":"Cell"},{"key":"2023101301523421400_B2","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1146\/annurev-pharmtox-010814-124316","article-title":"Genome integrity in aging: human syndromes, mouse models, and therapeutic options","volume":"56","author":"Vermeij","year":"2016","journal-title":"Annu. 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