RBFOX2 is critical for maintaining alternative polyadenylation patterns and mitochondrial health in rat myoblasts

Cell Rep. 2021 Nov 2;37(5):109910. doi: 10.1016/j.celrep.2021.109910.

Abstract

RBFOX2, which has a well-established role in alternative splicing, is linked to heart diseases. However, it is unclear whether RBFOX2 has other roles in RNA processing that can influence gene expression in muscle cells, contributing to heart disease. Here, we employ both 3'-end and nanopore cDNA sequencing to reveal a previously unrecognized role for RBFOX2 in maintaining alternative polyadenylation (APA) signatures in myoblasts. RBFOX2-mediated APA modulates mRNA levels and/or isoform expression of a collection of genes, including contractile and mitochondrial genes. Depletion of RBFOX2 adversely affects mitochondrial health in myoblasts, correlating with disrupted APA of mitochondrial gene Slc25a4. Mechanistically, RBFOX2 regulation of Slc25a4 APA is mediated through consensus RBFOX2 binding motifs near the distal polyadenylation site, enforcing the use of the proximal polyadenylation site. In sum, our results unveil a role for RBFOX2 in fine-tuning expression of mitochondrial and contractile genes via APA in myoblasts relevant to heart diseases.

Keywords: RBFOX2; Slc25a4; Tropomyosin 1; alternative polyadenylation; mitochondria; nanopore sequencing; poly(A) sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine Nucleotide Translocator 1 / genetics
  • Adenine Nucleotide Translocator 1 / metabolism
  • Animals
  • Gene Expression Regulation
  • HEK293 Cells
  • Humans
  • Mitochondria, Heart / genetics
  • Mitochondria, Heart / metabolism*
  • Mitochondria, Heart / ultrastructure
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism*
  • Muscle Proteins / genetics
  • Muscle Proteins / metabolism*
  • Myoblasts, Cardiac / metabolism*
  • Myoblasts, Cardiac / ultrastructure
  • Polyadenylation*
  • RNA Splicing Factors / genetics
  • RNA Splicing Factors / metabolism*
  • Rats
  • Tropomyosin / genetics
  • Tropomyosin / metabolism

Substances

  • Adenine Nucleotide Translocator 1
  • Mitochondrial Proteins
  • Muscle Proteins
  • RNA Splicing Factors
  • Rbfox2 protein, rat
  • Tpm1 protein, rat
  • Tropomyosin