Solution structure of the dyskeratosis congenita mutant P2b hairpin from human telomerase RNA
YNMG tetraloop formation by a dyskeratosis congenita mutation in human telomerase RNA.
Mutations linked to dyskeratosis congenita cause changes in the structural equilibrium in telomerase RNA
The wildtype P2b hairpin from human telomerase RNA
THE RNA WAS CHEMICALLY SYNTHESIZED using in vitro transcription with T7 RNA polymerase and a partially double-stranded DNA template. THE SEQUENCE OF THE RNA IS NATURALLY FOUND IN HOMO SAPIENS (HUMAN) who have a certain type of autosomal dominant dyskeratosis congenita.
CT-CE-HSQC was used for determining residual dipolar couplings
2D 15N-HMQC, 2D 15N-CPMG-NOESY, 2D JNN-HNN-COSY
3D 13C-NOESY HMQC, 2D 13C-HMQC,2D HCCH COSY, 3D HCCH TOCSY
Structures are based on 283 NOE-derived distance constraints, 86 dihedral angle restraints, 16 distance restraints from hydrogen bonds, and 14 H-C residual dipolar couplings.
1mM unlabeled RNA, 10mM sodium phosphate buffer, pH 6.3, 200mM KCl, 50uM EDTA, 0.2% sodium azide, 95% H2O,5% D2O
1mM 13C,15N-labeled RNA, 10mM sodium phosphate buffer, pH 6.3, 200mM KCl, 50uM EDTA, 0.2% sodium azide, 95% H2O,5% D2O
1mM unlabeled RNA, 10mM sodium phosphate buffer, pH 6.3, 200mM KCl, 50uM EDTA, 0.2% sodium azide, 100% D2O
1mM 13C,15N-labeled RNA, 10mM sodium phosphate buffer, pH 6.3, 200mM KCl, 50uM EDTA, 0.2% sodium azide, 100% D2O