Open AccessHeavy-atom labeling of RNA by PLOR for de novo crystallographic phasing
Author(s) -
Jason R. Stagno,
Ping Yü,
Marcin Dyba,
YunXing Wang,
Yu Liu
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0215555
Subject(s) - rna , phaser , riboswitch , aptamer , nucleotide , nucleic acid structure , crystallography , chemistry , computational biology , biology , non coding rna , physics , biochemistry , genetics , gene , optics
Due to the paucity of known RNA structures, experimental phasing is crucial for obtaining three-dimensional structures of RNAs by X-ray crystallography. Covalent attachment of heavy atoms to RNAs is one of the most useful strategies to facilitate phase determination. However, this approach is limited by the inefficiency or inability to synthesize large RNAs (>60 nucleotides) site-specifically labeled with heavy atoms using traditional methods. Here, we applied our recently reported method, PLOR (position-selective labeling of RNA) to incorporate 5-iodouridine at specific positions in the adenine riboswitch RNA aptamer domain, which was then used for crystallization and subsequent de novo SAD phasing. PLOR is a powerful tool to improve the efficiency of obtaining RNA structures de novo by X-ray crystallography.