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DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts.

Stokes, Matthew P. and Van Hatten, Ruth and Lindsay, Howard D. and Michael, W. Matthew (2002) DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts. Journal of Cell Biology, 158 (5). pp. 863-872. ISSN 0021-9525

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    Abstract

    Alkylating agents, such as methyl methanesulfonate (MMS), damage DNA and activate the DNA damage checkpoint. Although many of the checkpoint proteins that transduce damage signals have been identified and characterized, the mechanism that senses the damage and activates the checkpoint is not yet understood. To address this issue for alkylation damage, we have reconstituted the checkpoint response to MMS in Xenopus egg extracts. Using four different indicators for checkpoint activation (delay on entrance into mitosis, slowing of DNA replication, phosphorylation of the Chk1 protein, and physical association of the Rad17 checkpoint protein with damaged DNA), we report that MMS-induced checkpoint activation is dependent upon entrance into S phase. Additionally, we show that the replication of damaged double-stranded DNA, and not replication of damaged single-stranded DNA, is the molecular event that activates the checkpoint. Therefore, these data provide direct evidence that replication forks are an obligate intermediate in the activation of the DNA damage checkpoint.

    Item Type: Article
    Journal or Publication Title: Journal of Cell Biology
    Additional Information: © Rockefeller University Press
    Uncontrolled Keywords: cell cycle ; Rad17 ; DNA damage ; DNA replication ; S phase
    Subjects: Q Science > QH Natural history > QH301 Biology
    Departments: Faculty of Health and Medicine > Medicine
    ID Code: 9443
    Deposited By: Dr Howard Lindsay
    Deposited On: 10 Jun 2008 11:55
    Refereed?: Yes
    Published?: Published
    Last Modified: 26 Jul 2012 18:37
    Identification Number:
    URI: http://eprints.lancs.ac.uk/id/eprint/9443

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