5Y7Q

Crystal structure of paFAN1 bound to 2nt 5'flap DNA with gap

5Y7Q の概要

• エントリーDOI: 10.2210/pdb5y7q/pdb
• 分子名称: Fanconi-associated nuclease 1 homolog, DNA (5'-D(P*GP*TP*TP*GP*GP*GP*AP*TP*TP*G)-3'), DNA (5'-D(*TP*TP*CP*AP*CP*AP*CP*AP*TP*TP*CP*AP*A)-3'), ... (5 entities in total)
• 機能のキーワード: nuclease, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 81258.63

構造登録者

Jin, H.,Cho, Y. (登録日: 2017-08-17, 公開日: 2018-03-14, 最終更新日: 2023-11-22)

主引用文献

Jin, H.,Roy, U.,Lee, G.,Scharer, O.D.,Cho, Y.
Structural mechanism of DNA interstrand cross-link unhooking by the bacterial FAN1 nuclease.
J. Biol. Chem., 293:6482-6496, 2018

PubMed Abstract: DNA interstrand cross-links (ICLs) block the progress of the replication and transcription machineries and can weaken chromosomal stability, resulting in various diseases. FANCD2-FANCI-associated nuclease (FAN1) is a conserved structure-specific nuclease that unhooks DNA ICLs independently of the Fanconi anemia pathway. Recent structural studies have proposed two different mechanistic features for ICL unhooking by human FAN1: a specific basic pocket that recognizes the terminal phosphate of a 1-nucleotide (nt) 5' flap or FAN1 dimerization. Herein, we show that despite lacking these features, FAN1 (FAN1) cleaves substrates at ∼3-nt intervals and resolves ICLs. Crystal structures of FAN1 bound to various DNA substrates revealed that its conserved basic Arg/Lys patch comprising Arg-228 and Lys-260 recognizes phosphate groups near the 5' terminus of a DNA substrate with a 1-nt flap or a nick. Substitution of Lys-260 did not affect FAN1's initial endonuclease activity but significantly decreased its subsequent exonuclease activity and ICL unhooking. The Arg/Lys patch also interacted with phosphates at a 3-nt gap, and this interaction could drive movement of the scissile phosphates into the FAN1-active site. In human FAN1, the ICL-resolving activity was not affected by individual disruption of the Arg/Lys patch or basic pocket. However, simultaneous substitution of both FAN1 regions significantly reduced its ICL-resolving activity, suggesting that these two basic regions play a complementary role in ICL repair. On the basis of these findings, we propose a conserved role for two basic regions in FAN1 to guide ICL unhooking and to maintain genomic stability.

PubMed: 29514982
DOI: 10.1074/jbc.RA118.002171

実験手法

X-RAY DIFFRACTION (2.7 Å)