Abstract
γH2AX formation by phosphorylation of the histone variant H2AX is the key process in the repair of DNA lesions including those arising at fragile sites under replication stress. Here we demonstrate that H2AX is dynamically reorganized to preoccupy γH2AX hotspots on increased replication stress by activated cell proliferation and that H2AX is enriched in aphidicolin-induced replisome stalling sites in cycling cells. Interestingly, H2AX enrichment was particularly found in genomic regions that replicate in early S phase. High transcription activity, a hallmark of early replicating fragile sites, was a determinant of H2AX localization. Subtelomeric H2AX enrichment was also attributable to early replication and high gene density. In contrast, late replicating and infrequently transcribed regions, including common fragile sites and heterochromatin, lacked H2AX enrichment. In particular, heterochromatin was inaccessible to H2AX incorporation, maybe partly explaining the cause of mutation accumulation in cancer heterochromatin. Meanwhile, H2AX in actively dividing cells was intimately colocalized with INO80. INO80 silencing reduced H2AX levels, particularly at the INO80-enriched sites. Our findings suggest that active DNA replication is accompanied with the specific localization of H2AX and INO80 for efficient damage repair or replication-fork stabilization in actively transcribed regions.
Original language | English |
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Pages (from-to) | 1016-1025 |
Number of pages | 10 |
Journal | Nucleic Acids Research |
Volume | 42 |
Issue number | 2 |
DOIs | |
State | Published - 1 Jan 2014 |
Bibliographical note
Funding Information:KAIST Future Systems Healthcare Project from the Ministry of Science, ICT and Future Planning and by a grant from the National Research Foundation (NRF) of Korea [2012R1A1A1019094]. K.K. is a recipient of the Global PhD Fellowship of NRF. Computing facilities were supported by the NRF [2009-0086964] and the CHUNG Moon Soul Center of KAIST. Funding for open access charge: NRF [2012R1A1A1019094].