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Division of Labor between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment [BrdU-IP-ssSeq, ChIP-ssSeq and eSPAN analysis]

ABSTRACT: Division of Labor between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment [BrdU-IP-ssSeq, ChIP-ssSeq and eSPAN analysis]

PROVIDER: PRJNA574355 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10191123_1.fastq.gz	Fastqsanger.gz
	SRR10191123_2.fastq.gz	Fastqsanger.gz
	SRR10191124_1.fastq.gz	Fastqsanger.gz
	SRR10191124_2.fastq.gz	Fastqsanger.gz
	SRR10191125_1.fastq.gz	Fastqsanger.gz

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Budding yeast Wapl controls sister chromatid cohesion maintenance and the chromosome condensation status [BrdU-ChIP].

Project description:Cohesin acetylation by Eco1 during DNA replication establishes sister chromatid cohesion. We show that acetylation makes cohesin resistant to Wapl activity from S-phase until mitosis. Wapl turns out to be a key regulator of cohesin dynamics on chromosomes by controling cohesin maintenance following its establishment in S-phase and its role in chromosome condensation. The Affymetrix Saccharomyces cerevisiae Chip Tiling 1.0F Arrays were used to analyze the incorporation of BrdU in Saccharomyces cerevisiae in S-phase arrested cells.

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Division of Labor Between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment

Project description:Division of Labor Between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment

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Sister chromatid cohesion establishment by acetylation

Project description:Cohesion between sister chromatids depends on the chromosomal cohesin complex and allows the spindle apparatus in mitosis to recognize replicated chromosomes for segregation into daughter cells. Sister chromatid cohesion is established concomitant with DNA replication, and requires the essential Eco1 protein, a replication fork-associated acetyl transferase. The mechanism by which Eco1 establishes sister chromatid cohesion is not known. Here, we show that the cohesin subunit Smc3 is acetylated in an Eco1-dependent manner during S phase to establish sister chromatid cohesion. We isolated spontaneous suppressors of the thermosensitive eco1-1 allele in budding yeast, and identified the suppressor mutations from the hybridization pattern of genomic DNA on oligonucleotide tiling arrays. An acetylation mimicking mutation of a conserved lysine in Smc3 to asparagine (K113N) makes Eco1 dispensable for cell growth, indicating that Smc3 acetylation is Eco1’s only essential function. We identified a second set of eco1-1 suppressor mutations in the budding yeast ortholog of the cohesin regulator Wapl (Wpl1/Rad61). Wapl destabilizes cohesin on chromosomes, and Eco1-dependent Smc3 acetylation during S-phase might render cohesin resistant to Wapl. Our results clarify the role of Eco1 in the establishment of sister chromatid cohesion, and suggest that Eco1 modifies cohesin to stabilize an Eco1-independent cohesion establishment reaction.

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Division of Labor Between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment [Affymetrix ChIP-chip]

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SNW1 is required for sister chromatid cohesion

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