Analysis of Protein-RNA Complexes Involving a RNA Recognition Motif Engineered To Bind Hairpins with Seven- and Eight-Nucleotide Loops

作者：Blakeley Brett D; Shattuck Jenifer; Coates Melissa B; Tran Evelyn; Laird Offringa Ite A; McNaughton Brian R^*

来源：Biochemistry, 2013, 52(28): 4745-4747.

DOI：10.1021/bi400801q

U1A binds U1hpII, a hairpin RNA with a 10-nucleotide loop. A U1A mutant (Delta K50 Delta M51) binds U1hpII-derived hairpins with shorter loops, making it an interesting scaffold for engineering or evolving proteins that bind similarly sized disease-related hairpin RNAs. However, a more detailed understanding of complexes involving Delta K50 Delta M51 is likely a prerequisite to generating such proteins. Toward this end, we measured mutational effects for complexes involving U1A Delta K50 Delta M51 and U1hpII-derived hairpin RNAs with seven- or eight-nucleotide loops and identified contacts that are critical to the stabilization of these complexes. Our data provide valuable insight into sequence-selective recognition of seven- or eight-nucleotide loop hairpins by an engineered RNA binding protein.

• 出版日期2013-7-16