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. 2010 Jul 22;6(7):e1001032. doi: 10.1371/journal.pgen.1001032

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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(A) Genome-wide (ChIP-on-chip) distribution of γH2A during S phase. Schematics represent the three S. pombe chromosomes with key structural features (top). Tel- telomere; cen-centromere; MT-mating-type locus; rDNA-ribosomal DNA. Enrichment of γH2A is displayed as MAT scores (y-axis). Chromosome coordinates (x-axis, in megabases, (Mb)) downloaded from the S.pombe Genome Project (Sanger Center: www.sanger.ac.uk/Projects/S_pombe). (B) (left) γH2A formation occurs specifically during S phase. ChIP-qPCR timecourse analysis of γH2A enrichment at the indicated sites was performed by synchronizing cells using cdc25-22 block and release, and ChIP samples were collected every 30 min. Cell cycle progression was monitored by septation index. (right) Western blots comparing levels of γH2A in cdc25-22 wild type (Cds1+) and cdc25-22 cds1Δ cells released into 12 mM HU from G2 arrest. As an untreated control cdc25-22 wild type cells were released from G2 in the absence of HU. (C) Rad3 is the main kinase that phosphorylates H2A during unperturbed S phase. γH2AChIP-qPCR in wild type, rad3Δ, or tel1Δ cells synchronized as in (B), samples were collected in G2 and S phase. Primers: MT–5 kb from DSB in MT-locus; tel-subtelomere 1; cen-dh – centromere dh repeats; cnt – centromere 1 core; rDNA- 35S ribosomal DNA gene.