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J. Biol. Chem., Vol. 278, Issue 22, 19579-19582, May 30, 2003
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From the
Department of Oncology, Mayo Clinic and Foundation, Rochester, Minnesota 55905 and the ¶Division of Oncology, Columbia University, New York, New York 10032
53BP1 participates in the cellular response to DNA damage. Like many proteins involved in the DNA damage response, 53BP1 becomes hyperphosphorylated after radiation and colocalizes with phosphorylated H2AX in megabase regions surrounding the sites of DNA strand breaks. However, it is not yet clear whether the phosphorylation status of 53BP1 determines its localization or vice versa. In this study we mapped a region upstream of the 53BP1 C terminus that is required and sufficient for the recruitment of 53BP1 to these DNA break areas. In vitro assays revealed that this region binds to phosphorylated but not unphosphorylated H2AX. Moreover, using H2AX-deficient cells reconstituted with wild-type or a phosphorylation-deficient mutant of H2AX, we have shown that phosphorylation of H2AX at serine 140 is critical for efficient 53BP1 foci formation, implying that a direct interaction between 53BP1 and phosphorylated H2AX is required for the accumulation of 53BP1 at DNA break sites. On the other hand, radiation-induced phosphorylation of the 53BP1 N terminus by the ATM (ataxia-telangiectasia mutated) kinase is not essential for 53BP1 foci formation and takes place independently of 53BP1 redistribution. Thus, these two damage-induced events, hyperphosphorylation and relocalization of 53BP1, occur independently in the cell.
Received for publication, March 14, 2003 , and in revised form, April 15, 2003.
* This work was supported by grants from the National Institutes of Health, Breast Cancer Research Foundation, and Prospect Creek Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Supported by a postdoctoral fellowship from the Department of Defense breast cancer research program.
|| Recipient of a Department of Defense breast cancer career development award. To whom correspondence should be addressed. Tel.: 507-538-1545; Fax: 507-284-3906; E-mail: Chen.junjie{at}mayo.edu.
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