Qu, Meng, Rappas, Mathieu, Wardlaw, Christopher P, Garcia, Valerie, Ren, Jing-Yi, Day, Matthew, Carr, Antony M, Oliver, Antony, Du, Li-Lin and Pearl, Laurence (2013) Phosphorylation-dependent assembly and coordination of the DNA damage checkpoint apparatus by Rad4TopBP1. Molecular Cell, 51 (6). pp. 723-736. ISSN 1097-2765

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Abstract

The BRCT-domain protein Rad4(TopBP1) facilitates activation of the DNA damage checkpoint in Schizosaccharomyces pombe by physically coupling the Rad9-Rad1-Hus1 clamp, the Rad3(ATR) -Rad26(ATRIP) kinase complex, and the Crb2(53BP1) mediator. We have now determined crystal structures of the BRCT repeats of Rad4(TopBP1), revealing a distinctive domain architecture, and characterized their phosphorylation-dependent interactions with Rad9 and Crb2(53BP1). We identify a cluster of phosphorylation sites in the N-terminal region of Crb2(53BP1) that mediate interaction with Rad4(TopBP1) and reveal a hierarchical phosphorylation mechanism in which phosphorylation of Crb2(53BP1) residues Thr215 and Thr235 promotes phosphorylation of the noncanonical Thr187 site by scaffolding cyclin-dependent kinase (CDK) recruitment. Finally, we show that the simultaneous interaction of a single Rad4(TopBP1) molecule with both Thr187 phosphorylation sites in a Crb2(53BP1) dimer is essential for establishing the DNA damage checkpoint.

Item Type:	Article
Schools and Departments:	School of Life Sciences > Sussex Centre for Genome Damage and Stability
Subjects:	Q Science
Depositing User:	Deeptima Massey
Date Deposited:	04 Oct 2013 11:28
Last Modified:	02 Jul 2019 20:22
URI:	http://sro.sussex.ac.uk/id/eprint/46583

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