Science (New York, N.Y.) · 2023
In silico protein interaction screening uncovers DONSON's role in replication initiation.
СтатьяНазвание и аннотация: Перевод в очереди
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Lim Yang, Tamayo-Orrego Lukas, Schmid Ernst, Tarnauskaite Zygimante, Kochenova Olga V, Gruar Rhian, Muramatsu Sachiko, Lynch Luke, Schlie Aitana Verdu, Carroll Paula L, Chistol Gheorghe, Reijns Martin A M, Kanemaki Masato T, Jackson Andrew P, Walter Johannes C
Аннотация
CDC45-MCM2-7-GINS (CMG) helicase assembly is the central event in eukaryotic replication initiation. In yeast, a multi-subunit "pre-loading complex" (pre-LC) accompanies GINS to chromatin-bound MCM2-7, leading to CMG formation. Here, we report that DONSON, a metazoan protein mutated in microcephalic primordial dwarfism, is required for CMG assembly in vertebrates. 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 ε. 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.