Crystallization and preliminary X-ray analysis of human endonuclease 1 (APE1) in complex with an oligonucleotide containing a 5,6-dihydrouracil (DHU) or an alpha-anomeric 2'-deoxyadenosine (alphadA) modified base. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Acta crystallographica Section F : Structural biology communications [2014-...] Année : 2010

Crystallization and preliminary X-ray analysis of human endonuclease 1 (APE1) in complex with an oligonucleotide containing a 5,6-dihydrouracil (DHU) or an alpha-anomeric 2'-deoxyadenosine (alphadA) modified base.

Alexander A Ishchenko
Nikita A Kuznetsov
  • Fonction : Auteur
Murat Saparbaev
Solange Moréra

Résumé

The multifunctional human apurinic/apyrimidinic (AP) endonuclease 1 (APE1) is a key enzyme involved in both the base-excision repair (BER) and nucleotide-incision repair (NIR) pathways. In the NIR pathway, APE1 incises DNA 5' to a number of oxidatively damaged bases. APE1 was crystallized in the presence of a 15-mer DNA containing an oxidatively damaged base in a single central 5,6-dihydrouracil (DHU).T or alpha-anomeric 2'-deoxyadenosine (alphadA).T base pair. Diffraction data sets were collected to 2.2 and 2.7 A resolution from DNA-DHU-APE1 and DNA-alphadA-APE1 crystals, respectively. The crystals were isomorphous and contained one enzyme molecule in the asymmetric unit. Molecular replacement was performed and the initial electron-density maps revealed that in both complexes APE1 had crystallized with a degradation DNA product reduced to a 6-mer, suggesting that NIR and exonuclease reactions occurred prior to crystallization.

Dates et versions

hal-00512071 , version 1 (27-08-2010)

Identifiants

Citer

Pascal Retailleau, Alexander A Ishchenko, Nikita A Kuznetsov, Murat Saparbaev, Solange Moréra. Crystallization and preliminary X-ray analysis of human endonuclease 1 (APE1) in complex with an oligonucleotide containing a 5,6-dihydrouracil (DHU) or an alpha-anomeric 2'-deoxyadenosine (alphadA) modified base.. Acta crystallographica Section F : Structural biology communications [2014-..], 2010, 66 (Pt 7), pp.798-800. ⟨10.1107/S1744309110017021⟩. ⟨hal-00512071⟩
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