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Stop codon read-through of mammalian MTCH2 leading to an unstable isoform regulates mitochondrial membrane potential
- Source :
- The Journal of Biological Chemistry
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Stop codon read-through (SCR) is a process of continuation of translation beyond a stop codon. This phenomenon, which occurs only in certain mRNAs under specific conditions, leads to a longer isoform with properties different from that of the canonical isoform. MTCH2, which encodes a mitochondrial protein that regulates mitochondrial metabolism, was selected as a potential read-through candidate based on evolutionary conservation observed in the proximal region of its 3′ UTR. Here, we demonstrate translational read-through across two evolutionarily conserved, in-frame stop codons of MTCH2 using luminescence- and fluorescence-based assays, and by analyzing ribosome-profiling and mass spectrometry (MS) data. This phenomenon generates two isoforms, MTCH2x and MTCH2xx (single- and double-SCR products, respectively), in addition to the canonical isoform MTCH2, from the same mRNA. Our experiments revealed that a cis-acting 12-nucleotide sequence in the proximal 3′ UTR of MTCH2 is the necessary signal for SCR. Functional characterization showed that MTCH2 and MTCH2x were localized to mitochondria with a long t1/2 (>36 h). However, MTCH2xx was found predominantly in the cytoplasm. This mislocalization and its unique C terminus led to increased degradation, as shown by greatly reduced t1/2 (
- Subjects :
- translation control
0301 basic medicine
Gene isoform
Untranslated region
stop
mRNA
Protein degradation
Mitochondrion
Mitochondrial Membrane Transport Proteins
Biochemistry
Ribosome
Conserved sequence
03 medical and health sciences
mitochondrial membrane potential
stop codon
Animals
Humans
Protein Isoforms
RNA, Messenger
translational read-through
3' Untranslated Regions
Molecular Biology
Aorta
Membrane Potential, Mitochondrial
Messenger RNA
Base Sequence
030102 biochemistry & molecular biology
Chemistry
Cell Biology
Stop codon
MTCH2
Cell biology
mitochondria
HEK293 Cells
030104 developmental biology
ribosome
Protein Synthesis and Degradation
Protein Biosynthesis
protein degradation
Codon, Terminator
Cattle
Endothelium, Vascular
codon
CRISPR-Cas Systems
Ribosomes
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 295
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....145eff28cf6d96cc0d6dc303ed0b76d9