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• W2280239325 abstract "The initiation of DNA replication is a highly regulated process in eukaryotic cells, and central to the process of initiation is the assembly and activation of the replication fork helicase. The replication fork helicase is comprised of CMG (Cdc45, Mcm2–7, and GINS) in eukaryotic cells, and the mechanism underlying assembly of the CMG during S phase was studied in this article. We identified a point mutation of Sld3 that is specifically defective for Mcm3 and Mcm5 interaction (sld3-m10), and also identified a point mutation of Sld3 that is specifically defective for single-stranded DNA (ssDNA) interaction (sld3-m9). Expression of wild-type levels of sld3-m9 resulted in a severe DNA replication defect with no recruitment of GINS to Mcm2–7, whereas expression of wild-type levels of sld3-m10 resulted in a severe replication defect with no Cdc45 recruitment to Mcm2–7. We propose a model for Sld3-mediated control of replication initiation, wherein Sld3 manages the proper assembly of the CMG during S phase. We also find that the biochemical functions identified for Sld3 are conserved in human Treslin, suggesting that Treslin orchestrates assembly of the CMG in human cells. The initiation of DNA replication is a highly regulated process in eukaryotic cells, and central to the process of initiation is the assembly and activation of the replication fork helicase. The replication fork helicase is comprised of CMG (Cdc45, Mcm2–7, and GINS) in eukaryotic cells, and the mechanism underlying assembly of the CMG during S phase was studied in this article. We identified a point mutation of Sld3 that is specifically defective for Mcm3 and Mcm5 interaction (sld3-m10), and also identified a point mutation of Sld3 that is specifically defective for single-stranded DNA (ssDNA) interaction (sld3-m9). Expression of wild-type levels of sld3-m9 resulted in a severe DNA replication defect with no recruitment of GINS to Mcm2–7, whereas expression of wild-type levels of sld3-m10 resulted in a severe replication defect with no Cdc45 recruitment to Mcm2–7. We propose a model for Sld3-mediated control of replication initiation, wherein Sld3 manages the proper assembly of the CMG during S phase. We also find that the biochemical functions identified for Sld3 are conserved in human Treslin, suggesting that Treslin orchestrates assembly of the CMG in human cells. The replication initiation protein Sld3/Treslin orchestrates the assembly of the replication fork helicase during S phaseJournal of Biological ChemistryVol. 292Issue 24PreviewThis article has been withdrawn by the authors. The Dpb11 immunoblot from whole cell extracts in Fig. 4A and the Arp3 immunoblot from whole cell extracts in Fig. 4B were duplicated. Upon further investigation by the Journal, the Journal determined that the co-immunoprecipitation experiments shown in Fig. 4, A and B, were not performed appropriately. Full-Text PDF Open Access" @default.
• W2280239325 created "2016-06-24" @default.
• W2280239325 creator A5062800094 @default.
• W2280239325 creator A5066989891 @default.
• W2280239325 date "2015-11-01" @default.
• W2280239325 modified "2023-10-17" @default.
• W2280239325 title "The Replication Initiation Protein Sld3/Treslin Orchestrates the Assembly of the Replication Fork Helicase during S Phase" @default.
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• W2280239325 doi "https://doi.org/10.1074/jbc.m115.688424" @default.
• W2280239325 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5473237" @default.
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• W2280239325 hasPublicationYear "2015" @default.
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