Replication licensing regulated by a short linear motif within an intrinsically disordered region of origin recognition complex.
In eukaryotes, the origin recognition complex (ORC) faciliates the assembly of pre-replicative complex (pre-RC) at origin DNA for replication licensing. Here we show that the N-terminal intrinsically disordered region (IDR) of the yeast Orc2 subunit is crucial for this process. Removing a segment (residues 176-200) from Orc2-IDR or mutating a ... key isoleucine (194) significantly inhibits replication initiation across the genome. These Orc2-IDR mutants are capable of assembling the ORC-Cdc6-Cdt1-Mcm2-7 intermediate, which exhibits impaired ATP hydrolysis and fails to be convered into the subsequent Mcm2-7-ORC complex and pre-RC. These defects can be partially rescued by the Orc2-IDR peptide. Moreover, the phosphorylation of this Orc2-IDR region by S cyclin-dependent kinase blocks its binding to Mcm2-7 complex, causing a defective pre-RC assembly. Our findings provide important insights into the multifaceted roles of ORC in supporting origin licensing during the G1 phase and its regulation to restrict origin firing within the S phase.
Mesh Terms:
Amino Acid Motifs, Cell Cycle Proteins, DNA Replication, G1 Phase, Intrinsically Disordered Proteins, Mutation, Origin Recognition Complex, Phosphorylation, Protein Binding, Replication Origin, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins
Amino Acid Motifs, Cell Cycle Proteins, DNA Replication, G1 Phase, Intrinsically Disordered Proteins, Mutation, Origin Recognition Complex, Phosphorylation, Protein Binding, Replication Origin, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins
Nat Commun
Date: Sep. 13, 2024
PubMed ID: 39271725
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