CDK-dependent complex formation between replication proteins Dpb11, Sld2, Pol (epsilon}, and GINS in budding yeast.

By Sachiko Muramatsu, K. Hirai, Yon-Soo Tak, Y. Kamimura, Hiroyuki Araki. 2010

268 citations recorded by the source corpus.

Eukaryotic chromosomal DNA replication requires cyclin-dependent kinase (CDK) activity. CDK phosphorylates two yeast replication proteins, Sld3 and Sld2, both of which bind to Dpb11 when phosphorylated. These phosphorylation-dependent interactions are essential and are the minimal requirements for CDK-dependent activation of DNA replication. However, how these interactions activate DNA replication has not been elucidated. Here, we show that CDK promotes the formation of a newly identified fragile complex, the preloading complex (pre-LC) containing DNA polymerase epsilon (Pol epsilon), GINS, Sld2, and Dpb11. Formation of the pre-LC requires phosphorylation of Sld2 by CDK, but is independent of DNA replication, protein association with replication origins, and Dbf4-dependent Cdc7 kinase, which is also essential for the activation of DNA replication. We also demonstrate that Pol epsilon, GINS, Dpb11, and CDK-phosphorylated Sld2 form a complex in vitro. The genetic interactions between Pol epsilon, GINS, Sld2, and Dpb11 suggest further that they form an essential complex in cells. We propose that CDK regulates the initiation of DNA replication in budding yeast through formation of the pre-LC.

Subfields: Cellular and Molecular Neuroscience, Molecular Biology

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Citations in the corpus

The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells (2022, background): “According to a recent structural study (38), human POLE2 binds the GINS-MCM junction of the CMG helicase, which could make POLE2 essential for the CMG stability during some conformational changes, such as the polymerase switch step.”
DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 (2020, background): “Current models propose that a network of interactions between Sld3, Dpb11, Sld2, and GINS are required to form the CMG (Tanaka et al., 2007; Zegerman and Diffley, 2007; Muramatsu et al., 2010).”
Novel role of DONSON in CMG helicase assembly during vertebrate DNA replication initiation (2023, unknown): “Sld2 and Sld3 are the only two S-CDK targets that are essential for replication initiation (Tanaka et al, 2007; Zegerman & Diffley, 2007).”
Efficient Initiation of DNA Replication in Eukaryotes Requires Dpb11/TopBP1-GINS Interaction (2013, methodology): “BRCT1 and -2 and BRCT3 and -4 of Dpb11 bind to CDK-phosphorylated Sld3 and Sld2, respectively.”
DNSN-1 recruits GINS for CMG helicase assembly during DNA replication initiation in C. elegans (2023, unknown): “In this way, Dpb11 bridges phospho-Sld3 and Cdc45 on Mcm2-7 double hexamers to phospho-Sld2 that associates with GINS and DNA polymerase e (20, 21), leading to the formation of a preinitiation complex (step 4 in fig.”