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Series GSE240472 Query DataSets for GSE240472
Status Public on Jun 21, 2024
Title Mechanisms of MCM2-7 loading and initial DNA melting at near base-pair resolution
Organism Saccharomyces cerevisiae
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The loading and activation of the replicative helicase MCM2-7 are key events during the G1-S phase transition.In budding yeast, the origin recognition complex (ORC) binds to the conserved DNA elements A and B1 of the autonomously replicating sequence (ARS).This is followed by the consecutive loading of two MCM2-7 hetero-hexamers into a MCM2-7 double-hexamer(DH).In S-phase the MCM2-7DH is activated, resulting in two Cdc45-MCM-GINS(CMG) helicases that bidirectionally unwind DNA ahead of the replication fork.Here,we show that MCM2-7 helicase loading across the B1 element displaces ORC fromo rigins.This allows ORC binding and helicase loading at lower affinity binding sites and origins throughout the genome.Furthermore, we mapped the sites of initial DNA unwinding genome-wide and show that these sites appear near the N-terminal domains of the MCM2-7 double-hexamer in proximity of the B1 element.Finally, employing a chemical-biology approach, we establish that during helicase activation the Mcm2/5 interface acts as the DNA exit gate for single-stranded-DNA extrusion.Our work identifies that helicase loading follows a distributive mechanism, allowing for equal MCM2-7 loading across the genome and surprisingly finds that DNA unwinding initiates from the helicase N-terminal interface in proximity to the ARS B1 element.
 
Overall design ChIP-seq analysis of wt yeast cells, cdc6-1 and cdc46-1 mutants arrested in G2- and G1-phase to monitor pre-RC formation
 
Contributor(s) Reuter LM, Khadayate SP, Karimi MM, Speck C, Mossler A, Liebl K
Citation(s) 39181881
Submission date Aug 09, 2023
Last update date Sep 24, 2024
Contact name LMS Bioinformatics Core
E-mail(s) bioinformatics@lms.mrc.ac.uk
Organization name MRC London Institute of Medical Sciences
Department Bioinformatics Core
Street address Hammersmith Hospital Campus, Du Cane Road
City London
ZIP/Postal code W12 0NN
Country United Kingdom
 
Platforms (1)
GPL17342 Illumina HiSeq 2500 (Saccharomyces cerevisiae)
Samples (36)
GSM7698522 IP_Orc2-FLAG ChIP_Seq from wt cells arrested in G1 at the non-permissive temperature 37C, biol rep1
GSM7698523 IP_Orc2-FLAG ChIP_Seq from wt cells arrested in G1 at the non-permissive temperature 37C, biol rep2
GSM7698524 IP_Orc2-FLAG ChIP_Seq from wt cells arrested in G1 at the non-permissive temperature 37C, biol rep3
This SubSeries is part of SuperSeries:
GSE240779 MCM2-7 loading-dependent ORC release ensures genome-wide origin licensing
Relations
BioProject PRJNA1003856

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Supplementary file Size Download File type/resource
GSE240472_RAW.tar 1.5 Gb (http)(custom) TAR (of BW)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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