Molecular basis of XPF-ERCC1 targeting to SLX4-dependent DNA repair pathways

TL;DR

This paper reveals how the XPF-ERCC1 enzyme interacts with other proteins to repair DNA, using structural and functional studies.

Contribution

The study identifies key residues in XPF-ERCC1 that are crucial for its interaction with SLX4 and SLX4IP.

Findings

Point mutations in XPF-ERCC1 interfaces impair interactions with SLX4 or SLX4IP in human cells.

Disruption of the XPF-SLX4IP interface causes sensitivity to cis-platin.

The structure of the XPF-ERCC1-SLX4IP-SLX4330-555 complex with DNA is revealed.

Abstract

The preservation and faithful propagation of genetic information is essential for all life forms and depends on cellular pathways that enable replication, recombination, and repair of DNA. The multifunctional XPF-ERCC1 DNA endonuclease complex acts in several DNA repair pathways and interacts with numerous partner proteins and large DNA repair assemblies, including the nucleotide excision repair machinery and the SMX tri-endonuclease complex. Here, we report structures of XPF-ERCC1 in complex with the DNA repair factors SLX4 and SLX4IP, thereby identifying key residues responsible for direct interactions with XPF-ERCC1. When introduced into human cells, point mutations in these interfaces impair the interactions between XPF-ERCC1 and SLX4 or SLX4IP, and disruption of the XPF-SLX4IP interface leads to cis-platin sensitivity. Furthermore, our data reveal the structure of the human XPF-ERCC1-SLX4IP-SLX4330-555 complex with DNA bound at its active site, and they complete the structural characterisation of molecular interactions required to assemble the SMX complex.

Structure-specific endonucleases play an important role in several DNA repair pathways. Here the authors present structures of the endonuclease XPF-ERCC1 in complex with SLX4, SLX4IP, and DNA. Combined with functional analysis, these results provide insight into the mechanisms of XPF-ERCC1 recruitment and activation during DNA repair.

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