2MRF: NMR structure of the ubiquitin-binding zinc finger (UBZ) domain from human Rad18

Ubiquitin-mediated interactions are critical for the cellular DNA damage response (DDR). Therefore, many DDR-related proteins contain ubiquitin-binding domains, including ubiquitin-binding zinc fingers (UBZs). The majority of these UBZ domains belong to the C2H2 (type 3 Poleta-like) or C2HC (type 4 Rad18-like) family. We have used nuclear magnetic resonance (NMR) spectroscopy to characterize the binding to ubiquitin and determine the structure of the type 4 UBZ domain (UBZ4) from human Rad18, which is a key ubiquitin ligase in the DNA damage tolerance pathway responsible for monoubiquitination of the DNA sliding clamp PCNA. The Rad18-UBZ domain binds ubiquitin with micromolar affinity and adopts a beta1-beta2-alpha fold similar to the previously characterized type 3 UBZ domain (UBZ3) from the translesion synthesis DNA polymerase Poleta. However, despite nearly identical structures, a disparity in the location of binding-induced NMR chemical shift perturbations shows that the Rad18-UBZ4 and Poleta-UBZ3 domains bind ubiquitin in distinctly different modes. The Rad18-UBZ4 domain interacts with ubiquitin with the alpha-helix and strand beta1 as shown by the structure of the Rad18-UBZ domain-ubiquitin complex determined in this work, while the Poleta-UBZ3 domain exclusively utilizes the alpha-helix. Our findings suggest the existence of two classes of UBZ domains in DDR-related proteins with similar structures but unique ubiquitin binding properties and provide context for further study to establish the differential roles of these domains in the complex cellular response to DNA damage.
PDB ID: 2MRFDownload
MMDB ID: 123517
PDB Deposition Date: 2014/7/3
Updated in MMDB: 2014/10
Experimental Method:
solution nmr
Source Organism:
Similar Structures:
Biological Unit for 2MRF: monomeric; determined by author
Molecular Components in 2MRF
Label Count Molecule
Protein (1 molecule)
E3 Ubiquitin-protein Ligase Rad18(Gene symbol: RAD18)
Molecule annotation
Chemical (1 molecule)
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Citing MMDB