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| Status |
Public on May 16, 2023 |
| Title |
Isw1 contributes to cohesin association with centromeres and pericentromeric regions |
| Organisms |
Schizosaccharomyces pombe; Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Cohesin is a highly conserved, multiprotein complex whose canonical function is to hold sister chromatids together to ensure accurate chromosome segregation. Cohesin association with chromatin relies on the separate Scc2-Scc4 complex that enables cohesin ring opening and topological entrapment of sister DNAs. Cohesin loading at centromeres also requires the CTF19 kinetochore subcomplex that recruits Scc2-Scc4, ensuring efficient and targeted cohesin binding. After loading at centromeres, cohesin rapidly translocates to neighboring pericentromeric regions. In an attempt to better understand how sister chromatid cohesion is regulated, we performed a proteomic screen in budding yeast that identified the Isw1 chromatin remodeler as a cohesin binding partner. We found that while cohesin can associate with both Isw1-containing complexes, ISW1a and ISW1b, however only ISW1a localizes to centromeres. There, ISW1a utilizes its chromatin remodeling activity to allow efficient translocation of cohesin loaded through CTF19 at centromeres to pericentromeres. Consistently, lack of Isw1 results in cohesin and cohesin loader retainment at CENs and periCENs. Interestingly, lack of Isw1 resulted in changes in the nucleosomal landscape at the CENs and pericentromeric regions, suggesting that chromatin organization may be a crucial determinant of cohesin loading. Finally, we provide evidence that the role of ISW1a at the CENs is to restrict the activity of the RSC complex. Taken together, our results support the notion of a key role of chromatin environment for cohesin functions.
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| Overall design |
Calibrated Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for Scc1-9Pk and Rad21-6HA in Saccharomyces cerevisiae and Schizosaccharomyces pombe cells, respectively.
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| Contributor(s) |
Litwin I, Nowicka M, Maciaszczyk-Dziubińska E, Tomaszewska P, Markowska K, Wysocki R, Kramarz K |
| Citation(s) |
37486771 |
| |
| Submission date |
May 11, 2023 |
| Last update date |
Aug 16, 2023 |
| Contact name |
Ireneusz Bernard Litwin |
| E-mail(s) |
[email protected]
|
| Phone |
+48663491371
|
| Organization name |
University of Wroclaw
|
| Department |
Academic Excellence Incubator - Research Centre for DNA Repair and Replication
|
| Lab |
Karol Kramarz's lab
|
| Street address |
Kanonia 6/8
|
| City |
Wrocław |
| ZIP/Postal code |
50-328 |
| Country |
Poland |
| |
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| Platforms (1) |
| GPL29170 |
Illumina NovaSeq 6000 (Saccharomyces cerevisiae; Schizosaccharomyces pombe) |
|
| Samples (4)
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| GSM7326245 |
Calibrated Scc1 ChIP-seq for wild type IP |
| GSM7326246 |
Calibrated Scc1 ChIP-seq for wild type INPUT |
| GSM7326247 |
Calibrated Scc1 ChIP-seq for isw1Δ IP |
| GSM7326248 |
Calibrated Scc1 ChIP-seq for isw1Δ INPUT |
|
| Relations |
| BioProject |
PRJNA971597 |
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