Open AccessCrystallization and preliminary X‐ray analysis of a U2AF 65 variant in complex with a polypyrimidine‐tract analogue by use of protein engineering
Author(s) -
Sickmier E. Allen,
Frato Katherine E.,
Kielkopf Clara L.
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106012504
Subject(s) - crystallization , crystallography , x ray crystallography , materials science , phase (matter) , chemistry , physics , diffraction , optics , organic chemistry
The large subunit of the essential pre‐mRNA splicing factor U2 auxiliary factor (U2AF 65 ) binds the polypyrimidine tract near the 3′ splice site of pre‐mRNA introns and directs the association of the U2 small nuclear ribonucleoprotein particle (U2 snRNP) of the spliceosome with the pre‐mRNA. Protein engineering, in which the flexible linker region connecting tandem RNA‐recognition motifs (RRMs) within the U2AF 65 RNA‐binding domain was partially deleted, allowed successful crystallization of the protein–nucleic acid complex. Cocrystals of a U2AF 65 variant with a deoxyuridine dodecamer diffract X‐rays to 2.9 Å resolution and contain one complex per asymmetric unit.