title: The HhH domain of the human DNA repair protein XPF forms stable homodimers.
abstract: The human XPF-ERCC1 protein complex plays an essential role in nucleotide excision repair by catalysing positioned nicking of a DNA strand at the 5' side of the damage. We have recently solved the structure of the heterodimeric complex of the C-terminal domains of XPF and ERCC1 (Tripsianes et al., Structure 2005;13:1849-1858). We found that this complex comprises a pseudo twofold symmetry axis and that the helix-hairpin-helix motif of ERCC1 is required for DNA binding, whereas the corresponding domain of XPF is functioning as a scaffold for complex formation with ERCC1. Despite the functional importance of heterodimerization, the C-terminal domain of XPF can also form homodimers in vitro. We here compare the stabilities of homodimeric and heterodimeric complexes of the C-terminal domains of XPF and ERCC1. The higher stability of the XPF HhH complexes under various experimental conditions, determined using CD and NMR spectroscopy and mass spectrometry, is well explained by the structural differences that exist between the HhH domains of the two complexes. The XPF HhH homodimer has a larger interaction interface, aromatic stacking interactions, and additional hydrogen bond contacts as compared to the XPF/ERCC1 HhH complex, which accounts for its higher stability.
authors.affiliation: Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The Netherlands.
authors.forename: Devashish
authors.initials: D
authors.lastname: Das
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Tripsianes
authors.forename: Nicolaas G J
authors.initials: NG
authors.lastname: Jaspers
authors.forename: Jan H J
authors.initials: JH
authors.lastname: Hoeijmakers
authors.forename: Robert
authors.initials: R
authors.lastname: Kaptein
authors.forename: Rolf
authors.initials: R
authors.lastname: Boelens
authors.forename: Gert E
authors.initials: GE
authors.lastname: Folkers
mesh_terms: D024322:Amino Acids, Aromatic; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry*; D019281:Dimerization; D004720:Endonucleases / Q000737:chemistry*; D006801:Humans; D006860:Hydrogen Bonding; D046912:Multiprotein Complexes / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D024322:Amino Acids, Aromatic; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; C095888:xeroderma pigmentosum group F protein; C071447:ERCC1 protein, human; D004720:Endonucleases