Dose-Dependent Regulation of Alternative Splicing by MBNL Proteins Reveals Biomarkers for Myotonic Dystrophy.
Alternative splicing is a regulated process that results in expression of specific mRNA and protein isoforms. Alternative splicing factors determine the relative abundance of each isoform. Here we focus on MBNL1, a splicing factor misregulated in the disease myotonic dystrophy. By altering the concentration of MBNL1 in cells across a broad dynamic range, we show that different splicing events require different amounts of MBNL1 for half-maximal response, and respond more or less steeply to MBNL1. Motifs around MBNL1 exon 5 were studied to assess how cis-elements mediate the MBNL1 dose-dependent splicing response. A framework was developed to estimate MBNL concentration using splicing responses alone, validated in the cell-based model, and applied to myotonic dystrophy patient muscle. Using this framework, we evaluated the ability of individual and combinations of splicing events to predict functional MBNL concentration in human biopsies, as well as their performance as biomarkers to assay mild, moderate, and severe cases of DM.
Department of Chemistry and Biochemistry, Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.
Stacey D
http://orcid.org/0000-0003-3992-0798
SD
Wagner
Department of Chemistry and Biochemistry, Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.
Adam J
AJ
Struck
Department of Biology, Johns Hopkins University, Baltimore, Maryland, United States of America.
Riti
R
Gupta
Department of Biology, Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.
Dylan R
DR
Farnsworth
Department of Chemistry and Biochemistry, Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.
Amy E
AE
Mahady
Department of Neurology, University of Rochester Medical Center, Rochester, New York, United States of America.
Katy
K
Eichinger
Department of Neurology, University of Rochester Medical Center, Rochester, New York, United States of America.
Charles A
CA
Thornton
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.|Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.|Department of Biochemistry & Molecular Biology, Center for NeuroGenetics, University of Florida, Gainesville, Florida, United States of America.
Eric T
ET
Wang
Department of Chemistry and Biochemistry, Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.|Department of Biochemistry & Molecular Biology, Center for NeuroGenetics, University of Florida, Gainesville, Florida, United States of America.
J Andrew
JA
Berglund
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