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DNA duplex recognition activates Exo1 nuclease activity.

J. Biol. Chem.. 2019; 
LiYuxi,ShenJiangchuan,NiuHen
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Bacterial Expression System … 5 Page 6. Experimental procedures Expression and Purification of Exo1 from E. coli. The coding sequence of Exo1 (1-348) was re- designed through codon optimization by Genscript (sequence available upon request) in order to facilitate its expression in E. coli … Get A Quote

摘要

Exonuclease 1 (Exo1) is an evolutionarily conserved eukaryotic nuclease that plays a multifaceted role in maintaining genome stability. The biochemical attributes of Exo1 have been extensively characterized via conventional assays. However, the key step governing its activation remains elusive. Extending the previous finding that Exo1 can digest a randomly selected single-stranded DNA (ssDNA) but not a poly(dT) oligonucleotide and using purified recombinant Exo1 and nuclease and electrophoretic mobility shift assays, here we determined that DNA hairpins with a stem size of 4 bp or longer are able to activate Exo1-mediated digestion of ssDNA. We further provide evidence suggesting that Exo1 uses an evolution... More

关键词

DNA binding protein,DNA end resection,DNA endonuclease,DNA hairpin,DNA repair,DNA structure,Exo1,homologous recombination,replication protein A (