'AND' logic gates at work: Crystal structure of Rad53 bound to Dbf4 and Cdc7.
Forkhead-associated (FHA) domains are phosphopeptide recognition modules found in many signaling proteins. The Saccharomyces cerevisiae protein kinase Rad53 is a key regulator of the DNA damage checkpoint and uses its two FHA domains to interact with multiple binding partners during the checkpoint response. One of these binding partners is the Dbf4-dependent kinase (DDK), a heterodimer composed of the Cdc7 kinase and its regulatory subunit Dbf4. Binding of Rad53 to DDK, through its N-terminal FHA (FHA1) domain, ultimately inhibits DDK kinase activity, thereby preventing firing of late origins. We have previously found that the FHA1 domain of Rad53 binds simultaneously to Dbf4 and a phosphoepitope, suggesting that this domain functions as an 'AND' logic gate. Here, we present the crystal structures of the FHA1 domain of Rad53 bound to Dbf4, in the presence and absence of a Cdc7 phosphorylated peptide. Our results reveal how the FHA1 uses a canonical binding interface to recognize the Cdc7 phosphopeptide and a non-canonical interface to bind Dbf4. Based on these data we propose a mechanism to explain how Rad53 enhances the specificity of FHA1-mediated transient interactions.
Department of Biochemistry and Biomedical Sciences, ON, Canada.
Ahmad W
AW
Almawi
Department of Biochemistry and Biomedical Sciences, ON, Canada.
Lindsay A
LA
Matthews
Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Larasati
Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Polina
P
Myrox
Department of Chemistry and Chemical Biology, McMaster University, ON, Canada.
Stephen
S
Boulton
Department of Biochemistry, University of Toronto, Toronto, Canada.
Christine
C
Lai
Department of Biochemistry, University of Toronto, Toronto, Canada.
Trevor
T
Moraes
Department of Chemistry and Chemical Biology, McMaster University, ON, Canada.
Giuseppe
G
Melacini
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
Rodolfo
R
Ghirlando
Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Bernard P
BP
Duncker
Department of Biochemistry and Biomedical Sciences, ON, Canada.
Alba
A
Guarné
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