
Eukaryotic translation initiation factor 5A (eIF5A) is essential for alleviating ribosome stalling at the polyproline-rich region of the TIM50 mRNA, which encodes a key receptor subunit of the mitochondrial inner membrane translocase. Depletion of eIF5A leads to reduced Tim50 protein levels, compromised mitochondrial protein import, and activation of the mitochondrial protein import stress response.


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The efficient import of nuclear-encoded proteins into mitochondria is crucial for proper mitochondrial function. The conserved translation factor eIF5A is primarily known as an elongation factor which binds ribosomes to alleviate ribosome stalling at sequences encoding polyprolines or combinations of proline with glycine and charged amino acids. eIF5A is known to impact the mitochondrial function across a variety of species although the precise molecular mechanism underlying this impact remains unclear. We found that depletion of eIF5A in yeast drives reduced translation and levels of TCA cycle and oxidative phosphorylation proteins. We further found that loss of eIF5A leads to the accumulation of mitoprotein precursors in the cytosol as well as to the induction of a mitochondrial import stress response. Here we identify an essential polyproline-containing protein as a direct eIF5A target for translation: the mitochondrial inner membrane protein Tim50, which is the receptor sub-unit of the TIM23 translocase complex. We show how eIF5A directly controls mitochondrial protein import through the alleviation of ribosome stalling along TIM50 mRNA at the mitochondrial surface. Removal of the polyprolines from Tim50 rescues the mitochondrial import stress response, as well as the translation of oxidative phosphorylation reporter genes in an eIF5A loss of function. Overall, our findings elucidate how eIF5A impacts the mitochondrial function by reducing ribosome stalling and facilitating protein translation, thereby positively impacting the mitochondrial import process.

### Competing Interest Statement

The authors have declared no competing interest.

eIF5A controls mitoprotein import by relieving ribosome stalling at the TIM50 translocase mRNA

eukaryotic-translation-initiation-factor-5a-eif5a-regulates-mitochondrial-protein-import-by-alleviating-ribosome-stalling-at-the-tim50-translocase-mrna


Eukaryotic Translation Initiation Factor 5A (eIF5A) Regulates Mitochondrial Protein Import by Alleviating Ribosome Stalling at the TIM50 Translocase mRNA
