1EI4

B-DNA DODECAMER CGCGAAT(TLC)CGCG WITH INCORPORATED [3.3.0]BICYCLO-ARABINO-THYMINE-5'-PHOSPHATE

1EI4 の概要

• エントリーDOI: 10.2210/pdb1ei4/pdb
• 分子名称: DNA (5'-D(*CP*GP*CP*GP*AP*AP*(TLC)P*(TLC)P*CP*GP*CP*G)-3'), MAGNESIUM ION, SPERMINE, ... (4 entities in total)
• 機能のキーワード: modified b-dodecamer, [3.3.0]bicyclo-arabinonucleic acid, dna
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 7721.58

構造登録者

Minasov, G.,Teplova, M.,Nielsen, P.,Wengel, J.,Egli, M. (登録日: 2000-02-24, 公開日: 2000-03-27, 最終更新日: 2024-02-07)

主引用文献

Minasov, G.,Teplova, M.,Nielsen, P.,Wengel, J.,Egli, M.
Structural basis of cleavage by RNase H of hybrids of arabinonucleic acids and RNA.
Biochemistry, 39:3525-3532, 2000

PubMed Abstract: The origins of the substrate specificity of Escherichia coli RNase H1 (termed RNase H here), an enzyme that hydrolyzes the RNA strand of DNA-RNA hybrids, are not understood at present. Although the enzyme binds double-stranded RNA, no cleavage occurs with such duplexes [Lima, W. F., and Crooke, S. T. (1997) Biochemistry 36, 390]. Therefore, the hybrid substrates may not adopt a canonical A-form geometry. Furthermore, RNase H is exquisitely sensitive to chemical modification of the DNA strands in hybrid duplexes. This is particularly relevant to the RNase H-dependent pathway of antisense action. Thus, only very few of the modifications currently being evaluated as antisense therapeutics are tolerated by the enzyme, among them phosphorothioate DNA (PS-DNA). Recently, hybrids of RNA and arabinonucleic acid (ANA) as well as the 2'F-ANA analogue were shown to be substrates of RNase H [Damha, M. J., et al. (1998) J. Am. Chem. Soc. 120, 12976]. Using X-ray crystallography, we demonstrate here that ANA analogues, such as 2'F-ANA [Berger, I., et al. (1998) Nucleic Acids Res. 26, 2473] and [3.3.0]bicyclo-ANA (bc-ANA), may not be able to adopt sugar puckers that are compatible with pure A- or a B-form duplex geometries, but rather prefer the intermediate O4'-endo conformation. On the basis of the observed conformations of these ANA analogues in a DNA dodecamer duplex, we have modeled a duplex of an all-C3'-endo RNA strand and an all-O4'-endo 2'F-ANA strand. This duplex exhibits a minor groove width that is intermediate between that of A-form RNA and B-form DNA, a feature that may be exploited by the enzyme in differentiating between RNA duplexes and DNA-RNA hybrids. Therefore, the combination of the established structural and functional properties of ANA analogues helps settle existing controversies concerning the discrimination of substrates by RNase H. Knowlegde of the structure of an analogue that exhibits enhanced RNA affinity while not interfering with RNase H activity may prove helpful in the design of future antisense modifications.

PubMed: 10736151
DOI: 10.1021/bi992792j

実験手法

X-RAY DIFFRACTION (1.43 Å)