Journal Article

A Dbf4p BRCA1 C-Terminal-Like Domain Required for the Response to Replication Fork Arrest in Budding Yeast

Miller, Charles T, Gabrielse, Carrie, Fox, Catherine A, Weinreich, Michael, McConnell, Kristopher H, DeWard, Aaron

2006

Overview

Dbf4p is an essential regulatory subunit of the Cdc7p kinase required for the initiation of DNA replication. Cdc7p and Dbf4p orthologs have also been shown to function in the response to DNA damage. A previous Dbf4p multiple sequence alignment identified a conserved ∼40-residue N-terminal region with similarity to the BRCA1 C-terminal (BRCT) motif called “motif N.” BRCT motifs encode ∼100-amino-acid domains involved in the DNA damage response. We have identified an expanded and conserved ∼100-residue N-terminal region of Dbf4p that includes motif N but is capable of encoding a single BRCT-like domain. Dbf4p orthologs diverge from the BRCT motif at the C terminus but may encode a similar secondary structure in this region. We have therefore called this the BRCT and DBF4 similarity (BRDF) motif. The principal role of this Dbf4p motif was in the response to replication fork (RF) arrest; however, it was not required for cell cycle progression, activation of Cdc7p kinase activity, or interaction with the origin recognition complex (ORC) postulated to recruit Cdc7p–Dbf4p to origins. Rad53p likely directly phosphorylated Dbf4p in response to RF arrest and Dbf4p was required for Rad53p abundance. Rad53p and Dbf4p therefore cooperated to coordinate a robust cellular response to RF arrest.

• Publisher: Genetics Soc America,Genetics Society of America,Copyright © 2006 by the Genetics Society of America
• Subject: Amino Acid Motifs / Amino Acid Sequence / Amino acids / Breast cancer / Cell cycle / Cell Cycle Proteins - chemistry / Cell Cycle Proteins - genetics / Cell Cycle Proteins - metabolism / Deoxyribonucleic acid / DNA / DNA Damage / DNA polymerase / DNA repair / DNA Replication - genetics / DNA, Fungal - biosynthesis / DNA, Fungal - chemistry / DNA, Fungal - genetics / Genes, Fungal / Investigations / Models, Molecular / Molecular Sequence Data / Monoclonal antibodies / Point Mutation / Protein Conformation / Protein Structure, Tertiary / Protein Subunits / Protein-Serine-Threonine Kinases - chemistry / Protein-Serine-Threonine Kinases - genetics / Protein-Serine-Threonine Kinases - metabolism / Proteins / Saccharomyces cerevisiae / Saccharomyces cerevisiae - genetics / Saccharomyces cerevisiae - metabolism / Saccharomyces cerevisiae Proteins - chemistry / Saccharomyces cerevisiae Proteins - genetics / Saccharomyces cerevisiae Proteins - metabolism / Sequence Deletion / Sequence Homology, Amino Acid / Yeast / Yeasts