Dephosphorylation of the pre-initiation complex is critical for origin firing.
In eukaryotes, cyclin-dependent kinase (CDK) ensures that the genome is duplicated exactly once by inhibiting helicase loading factors before activating origin firing. CDK activates origin firing by phosphorylating two substrates, Sld2 and Sld3, forming a transient and limiting intermediate-the pre-initiation complex (pre-IC). Here, we show in the budding yeast Saccharomyces ... cerevisiae that the CDK phosphorylations of Sld3 and Sld2 are rapidly turned over during S phase by the PP2A and PP4 phosphatases. PP2ARts1 targets Sld3 specifically through an Rts1-interaction motif, and this targeted dephosphorylation is important for origin firing genome-wide, for formation of the pre-IC at origins and for ensuring that Sld3 is dephosphorylated in G1 phase. PP2ARts1 promotes replication in vitro, and we show that targeted Sld3 dephosphorylation is critical for viability. Together, these studies demonstrate that phosphatases enforce the correct ordering of replication factor phosphorylation and in addition to kinases are also key drivers of replication initiation.
Mesh Terms:
Cell Cycle Proteins, Cyclin-Dependent Kinases, DNA Replication, DNA-Binding Proteins, Phosphorylation, Replication Origin, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Saccharomycetales
Cell Cycle Proteins, Cyclin-Dependent Kinases, DNA Replication, DNA-Binding Proteins, Phosphorylation, Replication Origin, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Saccharomycetales
Mol Cell
Date: Jan. 05, 2023
PubMed ID: 36543171
View in: Pubmed Google Scholar
Download Curated Data For This Publication
246993
Switch View:
- Interactions 12