JBC Invitrogen Ultrasensitive Cytokine Assays

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Originally published In Press as doi:10.1074/jbc.M007665200 on October 20, 2000

J. Biol. Chem., Vol. 276, Issue 4, 2708-2718, January 26, 2001
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Negative Cell Cycle Regulation and DNA Damage-inducible Phosphorylation of the BRCT Protein 53BP1*

Zhenfang XiaDagger , Julio C. MoralesDagger , William G. Dunphy§, and Phillip B. CarpenterDagger ||**

From the Dagger  Department of Biochemistry and Molecular Biology, University of Texas Health Sciences Center, Houston, Texas 77030, the || Program in Cancer Biology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, and the § Howard Hughes Medical Institute and Division of Biology, California Institute of Technology, Pasadena, California 91125

In a screen designed to discover suppressors of mitotic catastrophe, we identified the Xenopus ortholog of 53BP1 (X53BP1), a BRCT protein previously identified in humans through its ability to bind the p53 tumor suppressor. X53BP1 transcripts are highly expressed in ovaries, and the protein interacts with Xp53 throughout the cell cycle in embryonic extracts. However, no interaction between X53BP1 and Xp53 can be detected in somatic cells, suggesting that the association between the two proteins may be developmentally regulated. X53BP1 is modified via phosphorylation in a DNA damage-dependent manner that correlates with the dispersal of X53BP1 into multiple foci throughout the nucleus in somatic cells. Thus, X53BP1 can be classified as a novel participant in the DNA damage response pathway. We demonstrate that X53BP1 and its human ortholog can serve as good substrates in vitro as well as in vivo for the ATM kinase. Collectively, our results reveal that 53BP1 plays an important role in the checkpoint response to DNA damage, possibly in collaboration with ATM.

* This work was supported in part by grants from the Ellison Medical Foundation and the Welch Foundation (to P. B. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF281071.

Investigator of the Howard Hughes Medical Institute.

** Junior Research Scholar of the Ellison Medical Foundation. To whom correspondence should be addressed. Tel.: 713-500-6032; Fax: 713-500-0652; E-mail: Phillip.B.Carpenter@uth.tmc.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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