A novel, non-canonical fha binding interface mediates the interaction between RAD53 and DBF4 [Protein Structure and Folding]

November 27th, 2013 by Matthews, L. A., Selvaratnam, R., Jones, D. R., Akimoto, M., McConkey, B. J., Melacini, G., Duncker, B. P., Guarne, A.

ForkHead Associated (FHA) and BRCA-1 C-terminal (BRCT) domains are overrepresented in DNA damage and replication stress response proteins. They primarily function as phosphoepitope-recognition modules, but can also mediate non-canonical interactions. The latter are rare and only a few have been studied at a molecular level. We have identified a crucial, non-canonical interaction between the N-terminal FHA1 domain of the checkpoint effector kinase Rad53 and the BRCT domain of the regulatory subunit of the Dbf4-dependent kinase, which is critical to suppress late origin firing and stabilize stalled forks during replication stress. The Rad53:Dbf4 interaction is phosphorylation-independent and involves a novel, non-canonical interface on the FHA1 domain. Mutations within this surface result in hypersensitivity to genotoxic stress. Importantly, this surface is not conserved in the FHA2 domain of Rad53, suggesting that the FHA domains of Rad53 gain specificity by engaging additional interaction interfaces beyond their phosphoepitope-binding site. In general, our results point to FHA domains functioning as complex logic gates rather than mere phosphoepitope targeting modules.