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Initiation-specific alleles of the Cdc45 helicase-activating protein
Name
journal.pone.0214426.pdf
Description
Published version
Size
1.98 MB
Format
Adobe PDF
Checksum (MD5)
c3d56a5f829bed8ba14880e0246bd1ba
Author(s)
Rios-Morales, Ramon Y
Chan, Sze Ham
Bell, Stephen P
Date Issued
2019
Journal
PLoS ONE
Publisher
Public Library of Science (PLoS)
Version
Final published version
Abstract
© 2019 Rios-Morales et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The committed step in DNA replication initiation is the activation of the Mcm2-7 replicative DNA helicase. Two activators, Cdc45 and GINS, associate with Mcm2-7 at origins of replication to form the CMG complex, which is the active eukaryotic replicative helicase. These activators function during both replication initiation and elongation, however, it remains unclear whether Cdc45 performs the same function(s) during both events. Here, we describe the genetic and biochemical characterization of seven Cdc45 mutations. Three of these mutations are temperature-sensitive lethal mutations in CDC45. Intriguingly, these mutants are defective for DNA replication initiation but not elongation. Consistent with an initiation defect, all three temperature-sensitive mutants are defective for CMG formation. Two of the lethal mutants are located within the RecJ-like domain of Cdc45 confirming the importance of this region for Cdc45 function. The remaining two lethal mutations localize to an intrinsically disordered region (IDR) of Cdc45 that is found in all eukaryotes. Despite the lethality of these IDR substitution mutants, Cdc45 lacking the IDR retains full function. Together, our data provide insights into the functional importance of Cdc45 domains and suggest that the requirements for Cdc45 function during DNA replication initiation are distinct from those involved in replication elongation.
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Creative Commons Attribution 4.0 International license
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DOI of Published Version
10.1371/journal.pone.0214426
