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Using AlphaFold to screen for protein-protein interactions followed by experimental validation, we show that DONSON scaffolds a vertebrate pre-LC containing GINS, TOPBP1, and DNA pol \u03b5. Our evidence suggests that DONSON docks the pre-LC onto MCM2-7, delivering GINS to its binding site in CMG. A patient-derived DONSON mutation compromises CMG assembly and recapitulates microcephalic dwarfism in mice. These results unify our understanding of eukaryotic replication initiation, implicate defective CMG assembly in microcephalic dwarfism, and illustrate how in silico protein-protein interaction screening accelerates mechanistic discovery.<\/jats:p>","DOI":"10.1126\/science.adi3448","type":"journal-article","created":{"date-parts":[[2023,8,17]],"date-time":"2023-08-17T13:56:36Z","timestamp":1692280596000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":130,"title":["In silico protein interaction screening uncovers DONSON\u2019s role in replication initiation"],"prefix":"10.1126","volume":"381","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6880-8418","authenticated-orcid":true,"given":"Yang","family":"Lim","sequence":"first","affiliation":[{"name":"Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7041-4848","authenticated-orcid":true,"given":"Lukas","family":"Tamayo-Orrego","sequence":"additional","affiliation":[{"name":"MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, EH4 2XU, UK."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4662-5298","authenticated-orcid":true,"given":"Ernst","family":"Schmid","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1815-1651","authenticated-orcid":true,"given":"Zygimante","family":"Tarnauskaite","sequence":"additional","affiliation":[{"name":"MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, EH4 2XU, UK."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga V.","family":"Kochenova","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA."},{"name":"Howard Hughes Medical Institute, Boston, MA 02115, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rhian","family":"Gruar","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sachiko","family":"Muramatsu","sequence":"additional","affiliation":[{"name":"Department of Chromosome Science, National Institute of Genetics, Research Organization of Information and Systems (ROIS), Mishima, Shizuoka 411-8540, Japan."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1948-2316","authenticated-orcid":true,"given":"Luke","family":"Lynch","sequence":"additional","affiliation":[{"name":"Biochemistry Department, Stanford School of Medicine, Stanford, CA 94305, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aitana Verdu","family":"Schlie","sequence":"additional","affiliation":[{"name":"MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, EH4 2XU, UK."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-0339-2559","authenticated-orcid":true,"given":"Paula L.","family":"Carroll","sequence":"additional","affiliation":[{"name":"MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, EH4 2XU, UK."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gheorghe","family":"Chistol","sequence":"additional","affiliation":[{"name":"Chemical and Systems Biology Department, Stanford School of Medicine, Stanford, CA 94305, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5048-2752","authenticated-orcid":true,"given":"Martin A. 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