title: Crystal structures of the structure-selective nuclease Mus81-Eme1 bound to flap DNA substrates.
abstract: The Mus81-Eme1 complex is a structure-selective endonuclease with a critical role in the resolution of recombination intermediates during DNA repair after interstrand cross-links, replication fork collapse, or double-strand breaks. To explain the molecular basis of 3' flap substrate recognition and cleavage mechanism by Mus81-Eme1, we determined crystal structures of human Mus81-Eme1 bound to various flap DNA substrates. Mus81-Eme1 undergoes gross substrate-induced conformational changes that reveal two key features: (i) a hydrophobic wedge of Mus81 that separates pre- and post-nick duplex DNA and (ii) a "5' end binding pocket" that hosts the 5' nicked end of post-nick DNA. These features are crucial for comprehensive protein-DNA interaction, sharp bending of the 3' flap DNA substrate, and incision strand placement at the active site. While Mus81-Eme1 unexpectedly shares several common features with members of the 5' flap nuclease family, the combined structural, biochemical, and biophysical analyses explain why Mus81-Eme1 preferentially cleaves 3' flap DNA substrates with 5' nicked ends.
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, South Korea.
authors.forename: Gwang Hyeon
authors.initials: GH
authors.lastname: Gwon
authors.forename: Aera
authors.initials: A
authors.lastname: Jo
authors.forename: Kyuwon
authors.initials: K
authors.lastname: Baek
authors.forename: Kyeong Sik
authors.initials: KS
authors.lastname: Jin
authors.forename: Yaoyao
authors.initials: Y
authors.lastname: Fu
authors.forename: Jong-Bong
authors.initials: JB
authors.lastname: Lee
authors.forename: Youngchang
authors.initials: Y
authors.lastname: Kim
authors.forename: Yunje
authors.initials: Y
authors.lastname: Cho
mesh_terms: D024506:5' Flanking Region; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry*; D053904:DNA Breaks, Single-Stranded; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004706:Endodeoxyribonucleases / Q000737:chemistry*; D004720:Endonucleases / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA; C507947:Eme1 protein, human; D004706:Endodeoxyribonucleases; D004720:Endonucleases; C498772:MUS81 protein, human
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