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| Status |
Public on Aug 22, 2013 |
| Title |
Telomeric repeats facilitate CENP-A incorporation via telomere binding proteins |
| Organism |
Schizosaccharomyces pombe |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
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| Summary |
The histone H3 variant, CENP-ACnp1, is normally assembled upon canonical centromeric sequences, but there is no apparent obligate coupling of sequence and assembly, suggesting that centromere location can be epigenetically determined. To explore the tolerances and constraints on CENP-ACnp1 deposition we investigated whether certain locations are favoured when additional CENP-ACnp1 is present in fission yeast cells. Our analyses show that additional CENP-ACnp1 accumulates within and close to heterochromatic centromeric outer repeats, and over regions adjacent to rDNA and telomeres. The use of minichromosome derivatives with unique DNA sequences internal to chromosome ends shows that telomeres are sufficient to direct CENP-ACnp1 deposition. However, chromosome ends are not required as CENP-ACnp1 deposition also occurs at telomere repeats inserted at an internal locus and correlates with the presence of H3K9 methylation near these repeats. The Ccq1 protein, which is known to bind telomere repeats and recruit telomerase, was found to be required to induce H3K9 methylation and thus promote the incorporation of CENP-A near telomere repeats. These analyses demonstrate that at non-centromeric chromosomal locations the presence of heterochromatin influences the sites at which CENP-A is incorporated into chromatin and thus, potentially the location of centromeres.
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| Overall design |
For CENP-A/Cnp1 chromatin immunoprecipitation: DNA immunoprecipitated with anti-Cnp1 serum using chromatin extracts from mutants and wild type control cells in biological duplicates normalized to input DNA from each strain.
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| Contributor(s) |
Castillo AG, Pidoux AL, Catania S, Durand-Dubief M, Choi ES, Hamilton G, Ekwall K, Allshire RC |
| Citation(s) |
23936074 |
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| Submission date |
Apr 26, 2013 |
| Last update date |
May 27, 2014 |
| Contact name |
Karl Ekwall |
| E-mail(s) |
karl.ekwall@ki.se
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| Phone |
+46 8 6089133
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| Organization name |
Karolinska Inst
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| Street address |
Alfred Nobels Alle 7
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| City |
Stockholm |
| ZIP/Postal code |
S-141 89 |
| Country |
Sweden |
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| Platforms (1) |
| GPL7715 |
[Sp20b_M] Affymetrix S. pombe Tiling 1.0FR Array |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA200313 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE46427_RAW.tar |
342.4 Mb |
(http)(custom) |
TAR (of BAR, CEL, TXT) |
| Processed data provided as supplementary file |
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