A Distal Auxiliary Element Facilitates Cleavage and Polyadenylation of Dux4 MRNA in the Pathogenic Haplotype of FSHD

Overview

Journal Hum Genet

Specialty Genetics

Date 2017 May 26

PMID 28540412

Citations 9

Authors

Natoya Peart

Eric J Wagner

Affiliations

Soon will be listed here.

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.

Citing Articles

The evolution of DUX4 gene regulation and its implication for facioscapulohumeral muscular dystrophy.

Jagannathan S Biochim Biophys Acta Mol Basis Dis. 2022; 1868(5):166367.

PMID: 35158020 PMC: 9173005. DOI: 10.1016/j.bbadis.2022.166367.

Systemic antisense therapeutics inhibiting DUX4 expression ameliorates FSHD-like pathology in an FSHD mouse model.

Lu-Nguyen N, Malerba A, Herath S, Dickson G, Popplewell L Hum Mol Genet. 2021; 30(15):1398-1412.

PMID: 33987655 PMC: 8283208. DOI: 10.1093/hmg/ddab136.

Current Therapeutic Approaches in FSHD.

Wang L, Tawil R J Neuromuscul Dis. 2021; 8(3):441-451.

PMID: 33579868 PMC: 8203219. DOI: 10.3233/JND-200554.

Transgenic mice expressing tunable levels of DUX4 develop characteristic facioscapulohumeral muscular dystrophy-like pathophysiology ranging in severity.

Jones T, Chew G, Barraza-Flores P, Schreier S, Ramirez M, Wuebbles R Skelet Muscle. 2020; 10(1):8.

PMID: 32278354 PMC: 7149937. DOI: 10.1186/s13395-020-00227-4.

Consequences of epigenetic derepression in facioscapulohumeral muscular dystrophy.

Greco A, Goossens R, van Engelen B, van der Maarel S Clin Genet. 2020; 97(6):799-814.

PMID: 32086799 PMC: 7318180. DOI: 10.1111/cge.13726.

References

Peart N, Wagner E . Gain-of-function reporters for analysis of mRNA 3'-end formation: Design and optimization. Biotechniques. 2016; 60(3):137-40. DOI: 10.2144/000114390. View

Zarkower D, Wickens M . A functionally redundant downstream sequence in SV40 late pre-mRNA is required for mRNA 3'-end formation and for assembly of a precleavage complex in vitro. J Biol Chem. 1988; 263(12):5780-8. View

Shi Y, Di Giammartino D, Taylor D, Sarkeshik A, Rice W, Yates 3rd J . Molecular architecture of the human pre-mRNA 3' processing complex. Mol Cell. 2009; 33(3):365-76. PMC: 2946185. DOI: 10.1016/j.molcel.2008.12.028. View

Barzaghi F, Passerini L, Bacchetta R . Immune dysregulation, polyendocrinopathy, enteropathy, x-linked syndrome: a paradigm of immunodeficiency with autoimmunity. Front Immunol. 2012; 3:211. PMC: 3459184. DOI: 10.3389/fimmu.2012.00211. View

Xiang K, Tong L, Manley J . Delineating the structural blueprint of the pre-mRNA 3'-end processing machinery. Mol Cell Biol. 2014; 34(11):1894-910. PMC: 4019069. DOI: 10.1128/MCB.00084-14. View

Lemmers R, Wohlgemuth M, Frants R, Padberg G, Morava E, van der Maarel S . Contractions of D4Z4 on 4qB subtelomeres do not cause facioscapulohumeral muscular dystrophy. Am J Hum Genet. 2004; 75(6):1124-30. PMC: 1182148. DOI: 10.1086/426035. View

Gehring N, Frede U, Neu-Yilik G, Hundsdoerfer P, Vetter B, Hentze M . Increased efficiency of mRNA 3' end formation: a new genetic mechanism contributing to hereditary thrombophilia. Nat Genet. 2001; 28(4):389-92. DOI: 10.1038/ng578. View

Chen F, MacDonald C, Wilusz J . Cleavage site determinants in the mammalian polyadenylation signal. Nucleic Acids Res. 1995; 23(14):2614-20. PMC: 307082. DOI: 10.1093/nar/23.14.2614. View

van Geel M, Dickson M, Beck A, Bolland D, Frants R, van der Maarel S . Genomic analysis of human chromosome 10q and 4q telomeres suggests a common origin. Genomics. 2002; 79(2):210-7. DOI: 10.1006/geno.2002.6690. View

10.

Ricci G, Scionti I, Sera F, Govi M, DAmico R, Frambolli I . Large scale genotype-phenotype analyses indicate that novel prognostic tools are required for families with facioscapulohumeral muscular dystrophy. Brain. 2013; 136(Pt 11):3408-17. PMC: 3808686. DOI: 10.1093/brain/awt226. View

11.

van Overveld P, Lemmers R, Sandkuijl L, Enthoven L, Winokur S, Bakels F . Hypomethylation of D4Z4 in 4q-linked and non-4q-linked facioscapulohumeral muscular dystrophy. Nat Genet. 2003; 35(4):315-7. DOI: 10.1038/ng1262. View

12.

Sheets M, Ogg S, Wickens M . Point mutations in AAUAAA and the poly (A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro. Nucleic Acids Res. 1990; 18(19):5799-805. PMC: 332317. DOI: 10.1093/nar/18.19.5799. View

13.

Shi Y, Manley J . The end of the message: multiple protein-RNA interactions define the mRNA polyadenylation site. Genes Dev. 2015; 29(9):889-97. PMC: 4421977. DOI: 10.1101/gad.261974.115. View

14.

Bennett C, Brunkow M, Ramsdell F, OBriant K, Zhu Q, Fuleihan R . A rare polyadenylation signal mutation of the FOXP3 gene (AAUAAA-->AAUGAA) leads to the IPEX syndrome. Immunogenetics. 2001; 53(6):435-9. DOI: 10.1007/s002510100358. View

15.

Bonano V, Oltean S, Garcia-Blanco M . A protocol for imaging alternative splicing regulation in vivo using fluorescence reporters in transgenic mice. Nat Protoc. 2007; 2(9):2166-81. DOI: 10.1038/nprot.2007.292. View

16.

Bosnakovski D, Xu Z, Gang E, Galindo C, Liu M, Simsek T . An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies. EMBO J. 2008; 27(20):2766-79. PMC: 2572182. DOI: 10.1038/emboj.2008.201. View

17.

Snider L, Geng L, Lemmers R, Kyba M, Ware C, Nelson A . Facioscapulohumeral dystrophy: incomplete suppression of a retrotransposed gene. PLoS Genet. 2010; 6(10):e1001181. PMC: 2965761. DOI: 10.1371/journal.pgen.1001181. View

18.

Marzluff W, Wagner E, Duronio R . Metabolism and regulation of canonical histone mRNAs: life without a poly(A) tail. Nat Rev Genet. 2008; 9(11):843-54. PMC: 2715827. DOI: 10.1038/nrg2438. View

19.

Scionti I, Greco F, Ricci G, Govi M, Arashiro P, Vercelli L . Large-scale population analysis challenges the current criteria for the molecular diagnosis of fascioscapulohumeral muscular dystrophy. Am J Hum Genet. 2012; 90(4):628-35. PMC: 3322229. DOI: 10.1016/j.ajhg.2012.02.019. View

20.

Tian B, Graber J . Signals for pre-mRNA cleavage and polyadenylation. Wiley Interdiscip Rev RNA. 2011; 3(3):385-96. PMC: 4451228. DOI: 10.1002/wrna.116. View