Low-density lipoprotein receptor-related protein 6 regulates alternative pre-mRNA splicing

J Cell Mol Med. 2018 Oct;22(10):4653-4663. doi: 10.1111/jcmm.13682. Epub 2018 Aug 1.

Abstract

Low-density lipoprotein receptor-related protein 6 (LRP6) serves as a Wnt coreceptor. Although Wnt/LRP6 signalling is best known for the β-catenin-dependent regulation of target genes in tissue development and homeostasis, emerging evidence demonstrates the biological aspects of LRP6 beyond a Wnt coreceptor. Whether LRP6 modulates tissue development in a Wnt/β-catenin signalling-independent manner remains unknown. Using a model of striated muscle development, we observed that LRP6 was almost undetectable in proliferating myoblasts, whereas its expression gradually increased in the nucleus of myodifferentiating cells. During myodifferentiation, LRP6 modulated the muscle-specific splicing of integrin-β1D and consequent myotube maturation independently of the β-catenin-dependent Wnt signalling. Furthermore, we identified that the carboxy-terminal serine-rich region in LRP6 bond to the adenine-rich sequence within alternative exon D (AED) of integrin-β1 pre-mRNA, and therefore, elicited AED inclusion when the spliceosome was recruited to the splice site. The interaction of LRP6 with the adenine-rich sequence was sufficient to overcome AED exclusion by a splicing repressor, polypyrimidine tract binding protein-1. Besides the integrin-β1, deep RNA sequencing in different types of cells revealed that the LRP6-mediated splicing regulation was widespread. Thus, our findings implicate LRP6 as a potential regulator for alternative pre-mRNA splicing.

Keywords: Wnt coreceptor; alternative splicing; integrin; lipoprotein receptor-related protein 6; myodifferentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing*
  • Animals
  • Animals, Newborn
  • Base Sequence
  • Cell Differentiation
  • Cell Line
  • Cell Nucleus / metabolism
  • Cell Proliferation
  • Cytosol / metabolism
  • Exons
  • Gene Expression Regulation, Developmental*
  • Heterogeneous-Nuclear Ribonucleoproteins / genetics
  • Heterogeneous-Nuclear Ribonucleoproteins / metabolism
  • Human Umbilical Vein Endothelial Cells / cytology
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Integrin beta1 / genetics
  • Integrin beta1 / metabolism
  • Low Density Lipoprotein Receptor-Related Protein-6 / genetics*
  • Low Density Lipoprotein Receptor-Related Protein-6 / metabolism
  • Mice
  • Muscle Development / genetics*
  • Muscle, Striated / cytology
  • Muscle, Striated / growth & development
  • Muscle, Striated / metabolism*
  • Myoblasts / cytology
  • Myoblasts / metabolism
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / metabolism
  • Polypyrimidine Tract-Binding Protein / genetics
  • Polypyrimidine Tract-Binding Protein / metabolism
  • Primary Cell Culture
  • RNA Precursors / genetics*
  • RNA Precursors / metabolism
  • Rats
  • Wnt Signaling Pathway
  • beta Catenin / genetics
  • beta Catenin / metabolism

Substances

  • CTNNB1 protein, mouse
  • Heterogeneous-Nuclear Ribonucleoproteins
  • Integrin beta1
  • Low Density Lipoprotein Receptor-Related Protein-6
  • Lrp6 protein, mouse
  • Ptbp1 protein, mouse
  • RNA Precursors
  • beta Catenin
  • Polypyrimidine Tract-Binding Protein

Associated data

  • GENBANK/ab37530
  • GENBANK/ab129115
  • GENBANK/ab133734
  • GENBANK/ab1835
  • GENBANK/ab180779
  • GENBANK/ab179471
  • GENBANK/ab8991
  • GENBANK/ab1791
  • GENBANK/ab123378
  • GENBANK/GSE102770