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Vol. 16, Issue 5, 2566-2576, May 2005

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Ionizing Radiation–dependent -H2AX Focus Formation Requires Ataxia Telangiectasia Mutated and Ataxia Telangiectasia Mutated and Rad3-related

Joanna D. Friesner ^*, Bo Liu , Kevin Culligan , and Anne B. Britt 

^* Genetics Graduate Group, University of California, Davis, Davis, CA 95616; Section of Plant Biology, University of California, Davis, Davis, CA 95616

Submitted October 14, 2004; Revised February 24, 2005; Accepted February 28, 2005
Monitoring Editor: Orna Cohen-Fix

The histone variant H2AX is rapidly phosphorylated at the sites of DNA double-strand breaks (DSBs). This phosphorylated H2AX (-H2AX) is involved in the retention of repair and signaling factor complexes at sites of DNA damage. The dependency of this phosphorylation on the various PI3K-related protein kinases (in mammals, ataxia telangiectasia mutated and Rad3-related [ATR], ataxia telangiectasia mutated [ATM], and DNA-PKCs) has been a subject of debate; it has been suggested that ATM is required for the induction of foci at DSBs, whereas ATR is involved in the recognition of stalled replication forks. In this study, using Arabidopsis as a model system, we investigated the ATR and ATM dependency of the formation of -H2AX foci in M-phase cells exposed to ionizing radiation (IR). We find that although the majority of these foci are ATM-dependent, 10% of IR-induced -H2AX foci require, instead, functional ATR. This indicates that even in the absence of DNA replication, a distinct subset of IR-induced damage is recognized by ATR. In addition, we find that in plants, -H2AX foci are induced at only one-third the rate observed in yeasts and mammals. This result may partly account for the relatively high radioresistance of plants versus yeast and mammals.

Address correspondence to: Anne B. Britt (abbritt{at}ucdavis.edu).

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