1D9H

Structural origins of the exonuclease resistance of a zwitterionic RNA

1D9H の概要

• エントリーDOI: 10.2210/pdb1d9h/pdb
• 分子名称: DNA/RNA (5'-D(*GP*CP*GP*TP*AP)-R(*(U31)P)-D(*AP*CP*GP*C)-3') (2 entities in total)
• 機能のキーワード: exonuclease resistance, a-dna, dna, rna
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 6208.15

構造登録者

Teplova, M.,Wallace, S.T.,Tereshko, V.,Minasov, G.,Simons, A.M.,Cook, P.D.,Manoharan, M.,Egli, M. (登録日: 1999-10-27, 公開日: 1999-12-02, 最終更新日: 2024-02-07)

主引用文献

Teplova, M.,Wallace, S.T.,Tereshko, V.,Minasov, G.,Symons, A.M.,Cook, P.D.,Manoharan, M.,Egli, M.
Structural origins of the exonuclease resistance of a zwitterionic RNA
Proc.Natl.Acad.Sci.USA, 96:14240-14245, 1999

PubMed Abstract: Nuclease resistance and RNA affinity are key criteria in the search for optimal antisense nucleic acid modifications, but the origins of the various levels of resistance to nuclease degradation conferred by chemical modification of DNA and RNA are currently not understood. The 2'-O-aminopropyl (AP)-RNA modification displays the highest nuclease resistance among all phosphodiester-based analogues and its RNA binding affinity surpasses that of phosphorothioate DNA by 1 degrees C per modified residue. We found that oligodeoxynucleotides containing AP-RNA residues at their 3' ends competitively inhibit the degradation of single-stranded DNA by the Escherichia coli Klenow fragment (KF) 3'-5' exonuclease and snake venom phosphodiesterase. To shed light on the origins of nuclease resistance brought about by the AP modification, we determined the crystal structure of an A-form DNA duplex with AP-RNA modifications at 1.6-A resolution. In addition, the crystal structures of complexes between short DNA fragments carrying AP-RNA modifications and wild-type KF were determined at resolutions between 2.2 and 3.0 A and compared with the structure of the complex between oligo(dT) and the D355A/E357A KF mutant. The structural models suggest that interference of the positively charged 2'-O-substituent with the metal ion binding site B of the exonuclease allows AP-RNA to effectively slow down degradation.

PubMed: 10588690
DOI: 10.1073/pnas.96.25.14240

実験手法

X-RAY DIFFRACTION (1.6 Å)