Browsing by Subject "C21orf57"

Mitochondrion is an essential organelle capable to produce high amounts of energy from oxidative phosphorylation (OXPHOS). The organelle counts on its own set of mitoribosomes and quality control (QC) factors to efficiently translate the OXPHOS components encoded by the mitochondrial DNA. Organelle dysfunction leads to development of mitochondrial diseases, which show a wide variety of symptoms and poor prognosis. ybeY is a highly conserved gene amongst prokaryotes that acts as factor for maturation and QC of bacterial ribosome. The ybeY-homolog C21orf57 is found remarkably conserved in eukaryotes, but its role in mammalian cells is completely unknown. In this study we characterised the mouse gene C21orf57 (mYBEY) in mammalian cells. mYBEY is transcribed at low levels in mouse tissues, where nervous tissue, skeletal and cardiac muscle show relatively higher amounts of mYBEY transcript. Endogenous mYbey protein could not be detected immunoblot due to its low expression level and/or rapid protein turnover. In parallel, mYBEY was cloned and successfully expressed in mammalian cells using transient and stable expression methodologies. We discovered that mYbey is imported into mitochondria and not covalently associated with mitoribosomes. Interestingly, protein translation stress and mitoribosome decay promoted by actinonin treatment is accompanied by a decrease in mYbey protein. In addition, depletion of mYbey using iRNA promotes the accumulation of mitoribosome proteins. We hypothesise a model in which mYbey is part of a QC mechanism for mitoribosome, and possibly involved with mitoribosome turnover. Although we successfully deleted mYBEY gene in mouse embryonic fibroblasts using the CRISPR/Cas9 approach, the isolation of viable mYBEY knock-out cells was not possible due to the possible deleterious and/or suppressive effects. In conclusion, we verified for the first time that mYbey is located within mammalian mitochondria and may be involved in a possible QC mechanism as a factor recruited for mitoribosome turnover. Further studies are necessary to fully elucidate the role of mYbey in the mitochondrial context.