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| Status |
Public on Jul 22, 2008 |
| Title |
A DNA polymerase α accessory protein, Mcl1, is required for propagation of centromere structures in fission yeast |
| Organism |
Schizosaccharomyces pombe |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
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| Summary |
Specialized chromatin exists at centromeres and must be precisely transmitted during DNA replication. The mechanisms involved in the propagation of these structures remain elusive. Fission yeast centromeres are composed of two chromatin domains: the central CENP-ACnp1 kinetochore domain and flanking heterochromatin domains. Here we show that fission yeast Mcl1, a DNA polymerase α (Polα) accessory protein, is critical for maintenance of centromeric chromatin. In a screen for mutants that alleviate both central domain and outer repeat silencing, we isolated several cos mutants, of which cos1 is allelic to mcl1. The mcl1-101 mutation causes reduced CENP-ACnp1 in the central domain and an aberrant increase in histone acetylation in both domains. These phenotypes are also observed in a mutant of swi7+, which encodes a catalytic subunit of Polα. Mcl1 forms S-phase-specific nuclear foci, which colocalize with those of PCNA and Polα. These results suggest that Mcl1 and Polα are required for propagation of centromere chromatin structures during DNA replication.
Keywords: ChIP-chip analysis, S. pombe
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| Overall design |
Antibodies used were α-acetyl-histone H4 antiserum (Upstate) and α-histone H4 pan antibody (Upstate). ChIP-on-chip was carried out using IVT amplification method. ChIP-chip data were comfirmed by real-time PCR.
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| Contributor(s) |
Natsume T, Tsutsui Y, Sutani T, Dunleavy EM, Pidoux AL, Iwasaki H, Shirahige K, Allshire RC, Yamano F |
| Citation(s) |
18493607 |
| |
| Submission date |
Apr 09, 2008 |
| Last update date |
Mar 19, 2012 |
| Contact name |
Katsuhiko Shirahige |
| E-mail(s) |
kshirahi@iam.u-tokyo.ac.jp
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| Phone |
+81-3-5842-0756
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| Fax |
+81-3-5842-0757
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| URL |
http://www.iam.u-tokyo.ac.jp/chromosomeinformatics/
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| Organization name |
The University of Tokyo
|
| Department |
Research Center for Epigenetic Disease
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| Lab |
Laboratory of Genome Structure and Function
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| Street address |
1-1-1 Yayoi
|
| City |
Bunkyo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
113-0032 |
| Country |
Japan |
| |
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| Platforms (1) |
| GPL3847 |
Affymetrix S. pombe Tiling 1.0FR Array |
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| Samples (4)
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| GSM280390 |
Histone H4 ChIP-chip data of Wild Type |
| GSM280391 |
Histone AcH4 ChIP-chip data of Wild Type |
| GSM280392 |
Histone H4 ChIP data of mcl1 deletion mutant |
| GSM280393 |
Histone Ac H4 ChIP fraction in mcl1 deletion mutant |
|
| Relations |
| BioProject |
PRJNA106985 |
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