Refinement of Trypanosoma U6 RNA Stem-Loop Structure Using 1- and 2-Bond Residual Dipolar Couplings
Authors | |
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Year of publication | 2004 |
Type | Article in Proceedings |
Conference | 19th NMR Valtice |
MU Faculty or unit | |
Citation | |
Field | Biochemistry |
Keywords | NMR; Residual dipolar couplings; RNA; structure; |
Description | The spliceosome is a complex of proteins and small nuclear RNAs (snRNAs) responsible for the removal of introns from pre-mRNA in eukaryotes. U6 snRNA has been demonstrated to be a vital element in splicing and its sequence is the most conserved of all snRNAs[1]. U6 snRNA from trypanosoma was our study. Determination of the solution structure of nucleic acid fragments is significantly improved if residual dipolar are measured. This data provides us additional long-range and local geometry restraints using a small degree of molecular alignment with the static magnetic field. We wanted to test polyethylenglycol (PEG) as a alignment medium for measuring the residual dipolar couplings (RDCs). PEG is a stable and unexpensive organic polymer that does not require a complex biochemical preparation like the most often used bacteriophage Pf1. In our study, applicability of PEG to several spin-state-selective E.COSY experiments was investigated. Measured RDCs were tested for internal consistency as refered in [2] in order to check the reliability of the values obtained in the PEG-aligned samples. |
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