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MBC in Press, published online ahead of print March 16, 2005
Mol. Biol. Cell 10.1091/mbc.E04-10-0890

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Submitted on October 14, 2004
Revised on February 24, 2005
Accepted on February 28, 2005

Ionizing Radiation-dependent {gamma}-H2AX Focus Formation Requires ATM and ATR

Joanna D. Friesner,* Bo Liu,{dagger} Kevin Culligan,{dagger} and Anne B. Britt{dagger}

*Genetics Graduate Group and {dagger}Section of Plant Biology, University of California, Davis, Davis, CA 95616

Monitoring Editor: Orna Cohen-Fix

The histone variant H2AX is rapidly phosphorylated at the sites of DNA double strand breaks (DSBs). This phosphorylated H2AX ({gamma}-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, ATR, 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, while 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 {gamma}-H2AX foci in M phase cells exposed to ionizing radiation (IR). We find that while the majority of these foci are ATM-dependent, ~10% of IR-induced {gamma}-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, {gamma}-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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