6D8M

Group I self-splicing intron P4-P6 domain mutant A125U/G126U

6D8M の概要

• エントリーDOI: 10.2210/pdb6d8m/pdb
• 分子名称: Group I self-splicing intron, MAGNESIUM ION (2 entities in total)
• 機能のキーワード: ribozyme, rna
• 由来する生物種: Tetrahymena thermophila
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 103195.92

構造登録者

Shoffner, G.M. (登録日: 2018-04-26, 公開日: 2018-07-04, 最終更新日: 2023-10-04)

主引用文献

Shoffner, G.M.,Wang, R.,Podell, E.,Cech, T.R.,Guo, F.
In Crystallo Selection to Establish New RNA Crystal Contacts.
Structure, 26:1275-, 2018

PubMed Abstract: Crystallography is a major technique for determining large RNA structures. Obtaining diffraction-quality crystals has been the bottleneck. Although several RNA crystallization methods have been developed, the field strongly needs additional approaches. Here we invented an in crystallo selection strategy for identifying mutations that enhance a target RNA's crystallizability. The strategy includes constructing an RNA pool containing random mutations, obtaining crystals, and amplifying the sequences enriched by crystallization. We demonstrated a proof-of-principle application to the P4-P6 domain from the Tetrahymena ribozyme. We further determined the structures of four selected mutants. All four establish new crystal lattice contacts while maintaining the native structure. Three mutants achieve this by relocating bulges and one by making a helix more flexible. In crystallo selection provides opportunities to improve crystals of RNAs or RNA-ligand complexes. Our results also suggest that mutants may be rationally designed for crystallization by "walking" a bulge along the RNA chain.

PubMed: 29910185
DOI: 10.1016/j.str.2018.05.005

実験手法

X-RAY DIFFRACTION (3.7 Å)