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A distal auxiliary element facilitates cleavage and polyadenylation of Dux4 mRNA in the pathogenic haplotype of FSHD

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Abstract

The degenerative muscle disorder facioscapulohumeral dystrophy (FSHD) is thought to be caused by the inappropriate expression of the Double Homeobox 4 (Dux4) protein in muscle cells leading to apoptosis. Expression of Dux4 in the major form of FSHD is a function of two contributing molecular changes: contractions in the D4Z4 microsatellite repeat region where Dux4 is located and an SNP present within a region downstream of the D4Z4. This SNP provides a functional, yet non-consensus polyadenylation signal (PAS) is used for the Dux4 mRNA 3′ end processing. Surprisingly, the sequences flanking the Dux4 PAS do not resemble a typical cleavage and polyadenylation landscape with no recognizable downstream sequence element and a suboptimal cleavage site. Here, we conducted a systematic analysis of the cis-acting elements that govern Dux4 cleavage and polyadenylation. Using a transcriptional read-through reporter, we determined that sequences downstream of the SNP located within the β-satellite region are critical for Dux4 cleavage and polyadenylation. We also demonstrate the feasibility of using antisense oligonucleotides to target these sequences as a means to reduce Dux4 expression. Our results underscore the complexity of the region immediately downstream of the D4Z4 and uncover a previously unknown function for the β-satellite region in Dux4 cleavage and polyadenylation.

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Acknowledgements

This work was supported by the Muscular Dystrophy Foundation (MDA 202141), Friends of FSH Research, and a Schissler Foundation Fellowship for Translational Studies of Common Human Diseases.

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  1. Natoya Peart

    Present address: Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch at Galveston, Galveston, USA

    Natoya Peart & Eric J. Wagner

  2. Graduate Program in Biochemistry and Molecular Biology, The University of Texas Graduate School of Biomedical Sciences, Houston, TX, USA

    Natoya Peart

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  1. Natoya Peart
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NP performed the experiments and data analyses, and wrote the manuscript. EJW conceived the project and contributed to the design of the experiments, and manuscript writing.

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Correspondence to Eric J. Wagner.

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Peart, N., Wagner, E.J. A distal auxiliary element facilitates cleavage and polyadenylation of Dux4 mRNA in the pathogenic haplotype of FSHD. Hum Genet 136, 1291–1301 (2017). https://doi.org/10.1007/s00439-017-1813-8

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  • DOI: https://doi.org/10.1007/s00439-017-1813-8

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