RAD54 N-terminal domain is a DNA sensor that couples ATP hydrolysis with branch migration of Holliday junctions.
Publication Title
Nat Commun
Document Type
Article
Publication Date
1-2-2018
Keywords
Adenosine Triphosphatases; Amino Acid Sequence; Animals; Binding Sites; Cell Line; DNA Helicases; DNA Repair; DNA, Cruciform; Humans; Hydrolysis; Nuclear Proteins; Nucleic Acid Conformation; Phosphorylation; Protein Multimerization; Recombination, Genetic; Sequence Homology, Amino Acid; Sf9 Cells; Spodoptera
Abstract
In eukaryotes, RAD54 catalyzes branch migration (BM) of Holliday junctions, a basic process during DNA repair, replication, and recombination. RAD54 also stimulates RAD51 recombinase and has other activities. Here, we investigate the structural determinants for different RAD54 activities. We find that the RAD54 N-terminal domain (NTD) is responsible for initiation of BM through two coupled, but distinct steps; specific binding to Holliday junctions and RAD54 oligomerization. Furthermore, we find that the RAD54 oligomeric state can be controlled by NTD phosphorylation at S49, a CDK2 consensus site, which inhibits RAD54 oligomerization and, consequently, BM. Importantly, the effect of phosphorylation on RAD54 oligomerization is specific for BM, as it does not affect stimulation of RAD51 recombinase by RAD54. Thus, the transition of the oligomeric states provides an important control of the biological functions of RAD54 and, likely, other multifunctional proteins.
Specialty/Research Institute
Institute for Systems Biology
