title: Pseudouridine Modification Inhibits Muscleblind-like 1 (MBNL1) Binding to CCUG Repeats and Minimally Structured RNA through Reduced RNA Flexibility.
abstract: Myotonic dystrophy type 2 is a genetic neuromuscular disease caused by the expression of expanded CCUG repeat RNAs from the non-coding region of theCCHC-type zinc fingernucleic acid-bindingprotein (CNBP) gene. These CCUG repeats bind and sequester a family of RNA-binding proteins known as Muscleblind-like 1, 2, and 3 (MBNL1, MBNL2, and MBNL3), and sequestration plays a significant role in pathogenicity. MBNL proteins are alternative splicing regulators that bind to the consensus RNA sequence YGCY (Y = pyrimidine). This consensus sequence is found in the toxic RNAs (CCUG repeats) and in cellular RNA substrates that MBNL proteins have been shown to bind. Replacing the uridine in CCUG repeats with pseudouridine (Ψ) resulted in a modest reduction of MBNL1 binding. Interestingly, Ψ modification of a minimally structured RNA containing YGCY motifs resulted in more robust inhibition of MBNL1 binding. The different levels of inhibition between CCUG repeat and minimally structured RNA binding appear to be due to the ability to modify both pyrimidines in the YGCY motif, which is not possible in the CCUG repeats. Molecular dynamic studies of unmodified and pseudouridylated minimally structured RNAs suggest that reducing the flexibility of the minimally structured RNA leads to reduced binding by MBNL1.
authors.affiliation: From the Institute of Molecular Biology, Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403 and.
authors.forename: Elaine
authors.initials: E
authors.lastname: deLorimier
authors.affiliation: From the Institute of Molecular Biology, Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403 and.
authors.forename: Melissa N
authors.initials: MN
authors.lastname: Hinman
authors.affiliation: From the Institute of Molecular Biology, Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403 and.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Copperman
authors.affiliation: the Center for NeuroGenetics, Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida 32610-3010.
authors.forename: Kausiki
authors.initials: K
authors.lastname: Datta
authors.affiliation: From the Institute of Molecular Biology, Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403 and.
authors.forename: Marina
authors.initials: M
authors.lastname: Guenza
authors.affiliation: From the Institute of Molecular Biology, Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403 and aberglund@ufl.edu.|the Center for NeuroGenetics, Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida 32610-3010.
authors.forename: J Andrew
authors.initials: JA
authors.lastname: Berglund
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D006801:Humans; D007438:Introns; D056004:Molecular Dynamics Simulation; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D011560:Pseudouridine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*
chemical_list: C493663:MBNL1 protein, human; D016601:RNA-Binding Proteins; D011560:Pseudouridine; D012313:RNA
keywords: RNA modification; RNA structure; RNA-binding protein; alternative splicing; molecular dynamics