In Crystallo Selection to Establish New RNA Crystal Contacts.

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 ach

SUBMITTER: Shoffner GM 

PROVIDER: S-EPMC6125213 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature