5K97

Flap endonuclease 1 (FEN1) D233N with cleaved product fragment and Sm3+

5K97 の概要

• エントリーDOI: 10.2210/pdb5k97/pdb
• 関連するPDBエントリー: 5K96
• 分子名称: Flap endonuclease 1, DNA (5'-D(*AP*CP*TP*CP*TP*GP*CP*CP*TP*CP*AP*AP*GP*AP*CP*GP*GP*T)-3'), DNA (5'-D(P*TP*GP*AP*GP*GP*CP*AP*GP*AP*GP*T)-3'), ... (9 entities in total)
• 機能のキーワード: metalloprotein, replication, dna damage, dna repair, base excision repair, protein-dna, 5' nuclease, fen, product, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 52266.46

構造登録者

Tsutakawa, S.E.,Arvai, A.S.,Tainer, J.A. (登録日: 2016-05-31, 公開日: 2017-06-28, 最終更新日: 2024-02-28)

主引用文献

Tsutakawa, S.E.,Thompson, M.J.,Arvai, A.S.,Neil, A.J.,Shaw, S.J.,Algasaier, S.I.,Kim, J.C.,Finger, L.D.,Jardine, E.,Gotham, V.J.B.,Sarker, A.H.,Her, M.Z.,Rashid, F.,Hamdan, S.M.,Mirkin, S.M.,Grasby, J.A.,Tainer, J.A.
Phosphate steering by Flap Endonuclease 1 promotes 5'-flap specificity and incision to prevent genome instability.
Nat Commun, 8:15855-15855, 2017

PubMed Abstract: DNA replication and repair enzyme Flap Endonuclease 1 (FEN1) is vital for genome integrity, and FEN1 mutations arise in multiple cancers. FEN1 precisely cleaves single-stranded (ss) 5'-flaps one nucleotide into duplex (ds) DNA. Yet, how FEN1 selects for but does not incise the ss 5'-flap was enigmatic. Here we combine crystallographic, biochemical and genetic analyses to show that two dsDNA binding sites set the 5'polarity and to reveal unexpected control of the DNA phosphodiester backbone by electrostatic interactions. Via 'phosphate steering', basic residues energetically steer an inverted ss 5'-flap through a gateway over FEN1's active site and shift dsDNA for catalysis. Mutations of these residues cause an 18,000-fold reduction in catalytic rate in vitro and large-scale trinucleotide (GAA) repeat expansions in vivo, implying failed phosphate-steering promotes an unanticipated lagging-strand template-switch mechanism during replication. Thus, phosphate steering is an unappreciated FEN1 function that enforces 5'-flap specificity and catalysis, preventing genomic instability.

PubMed: 28653660
DOI: 10.1038/ncomms15855

実験手法

X-RAY DIFFRACTION (2.102 Å)