SANS (USH1G) regulates pre-mRNA splicing by mediating the intra-nuclear transfer of tri-snRNP complexes

Nucleic Acids Res. 2021 Jun 4;49(10):5845-5866. doi: 10.1093/nar/gkab386.

Abstract

Splicing is catalyzed by the spliceosome, a compositionally dynamic complex assembled stepwise on pre-mRNA. We reveal links between splicing machinery components and the intrinsically disordered ciliopathy protein SANS. Pathogenic mutations in SANS/USH1G lead to Usher syndrome-the most common cause of deaf-blindness. Previously, SANS was shown to function only in the cytosol and primary cilia. Here, we have uncovered molecular links between SANS and pre-mRNA splicing catalyzed by the spliceosome in the nucleus. We show that SANS is found in Cajal bodies and nuclear speckles, where it interacts with components of spliceosomal sub-complexes such as SF3B1 and the large splicing cofactor SON but also with PRPFs and snRNAs related to the tri-snRNP complex. SANS is required for the transfer of tri-snRNPs between Cajal bodies and nuclear speckles for spliceosome assembly and may also participate in snRNP recycling back to Cajal bodies. SANS depletion alters the kinetics of spliceosome assembly, leading to accumulation of complex A. SANS deficiency and USH1G pathogenic mutations affects splicing of genes related to cell proliferation and human Usher syndrome. Thus, we provide the first evidence that splicing dysregulation may participate in the pathophysiology of Usher syndrome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing / genetics*
  • Cell Nucleus / metabolism
  • Cell Proliferation / genetics
  • Coiled Bodies / metabolism
  • DNA-Binding Proteins / metabolism
  • Eye Proteins / metabolism
  • Gene Knockdown Techniques
  • HEK293 Cells
  • Humans
  • Immunohistochemistry
  • In Situ Hybridization, Fluorescence
  • Mass Spectrometry
  • Microscopy, Electron, Transmission
  • Minor Histocompatibility Antigens / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Phosphoproteins / metabolism
  • Proteomics
  • RNA Precursors / genetics
  • RNA Precursors / metabolism*
  • RNA Splicing Factors / metabolism
  • RNA, Small Nuclear / genetics
  • RNA, Small Nuclear / metabolism
  • Ribonucleoprotein, U4-U6 Small Nuclear / metabolism
  • Ribonucleoproteins, Small Nuclear / genetics
  • Ribonucleoproteins, Small Nuclear / metabolism*
  • Spliceosomes / genetics
  • Spliceosomes / metabolism*
  • Transcription Factors / metabolism
  • Usher Syndromes / genetics
  • Usher Syndromes / metabolism*

Substances

  • DNA-Binding Proteins
  • Eye Proteins
  • Minor Histocompatibility Antigens
  • Nerve Tissue Proteins
  • PRPF31 protein, human
  • PRPF6 protein, human
  • Phosphoproteins
  • RNA Precursors
  • RNA Splicing Factors
  • RNA, Small Nuclear
  • Ribonucleoprotein, U4-U6 Small Nuclear
  • Ribonucleoproteins, Small Nuclear
  • SF3B1 protein, human
  • SON protein, human
  • Transcription Factors
  • USH1G protein, human