APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN  ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS

Mitochondrial translation is a highly specialized process of synthesizing mitochondrically encoded proteins, mainly the components of the oxidative phosphorylation system. It involves four key stages: initiation, elongation, termination, and recycling of mitochondrial ribosomes. Each of these stages is regulated by a specific set of translation factors, most of which are encoded by the nuclear genome and imported into mitochondria. The termination of mitochondrial translation in yeast () is carried out by the MRF1 release factor. This nuclear-encoded factor is crucial for ensuring accurate protein synthesis within the organelle, as it recognizes stop codons and facilitates the release of completed polypeptide chains from the ribosome. In addition to its main function, MRF1 participates in maintaining mitochondrial genome stability. The aim of this study was to investigate the capacity of human homologs, hMTRF1, hMTRF1A, and mitoribosome rescue factors hMTRFR and hMRPL58, to compensate for the absence of the yeast mitochondrial translation termination factor MRF1 in cells. The results obtained suggest that human orthologs of MRF1, such as hMTRF1 and hMTRF1A, can contribute to maintaining the integrity of yeast mitochondrial genome. However, they do not fully make up for the absence of MRF1, as they do not restore normal respiration of the mutant yeast strains.

Keywords

пекарские дрожжи митохондрии трансляция биосинтез белка терминация митохондриальная ДНК гуманизация

Date of publication

20.03.2026

Year of publication

2026

Number of purchasers

Views

References

1. Wang, F., Zhang, D., Zhang, D., Li, P., and Gao, Y. (2021) Mitochondrial protein translation: emerging roles and clinical significance in disease, Front. Cell Dev. Biol., 9, 675475, https://doi.org/10.3389/fcell.2021.675465.
2. Kummer, E., and Ban, N. (2021) Mechanisms and regulation of protein synthesis in mitochondria, Nat. Rev. Mol. Cell Biol., 22, 307-325, https://doi.org/10.1038/s41580-021-00332-2.
3. Pel, H. J., Maat, C., Rep, M., and Grivell, L. A. (1992) The yeast nuclear gene MRF1 encodes a mitochondrial peptide chain release factor and cures several mitochondrial RNA splicing defects, Nucleic Acids Res., 20, 6339-6346, https://doi.org/10.1093/nar/20.23.6339.
4. Towpik, J., Chacinska, A., Ciesla, M., Ginalski, K., and Boguta, M. (2004) Mutations in the yeast mrf1 gene encoding mitochondrial release factor inhibit translation on mitochondrial ribosomes, J. Biol. Chem., 279, 14096-14103, https://doi.org/10.1074/jbc.M312856200.
5. Krüger, A., Kovalchuk, D., Shiriaev, D., and Rorbach, J. (2024) Decoding the enigma: translation termination in human mitochondria, Hum. Mol. Genet., 33, R42-R46, https://doi.org/10.1093/hmg/ddae032.
6. Merz, S., and Westermann, B. (2009) Genome-wide deletion mutant analysis reveals genes required for respiratory growth, mitochondrial genome maintenance and mitochondrial protein synthesis in Saccharomyces cerevisiae, Genome Biol., 10, R95, https://doi.org/10.1186/gb-2009-10-9-r95.
7. Laurent, J. M., Garge, R. K., Teufel, A. I., Wilke, C. O., Kachroo, A. H., and Marcotte, E. M. (2020) Humanization of yeast genes with multiple human orthologs reveals functional divergence between paralogs, PLoS Biol., 18, e3000627, https://doi.org/10.1371/journal.pbio.3000627.
8. Hamza, A., Driessen, M. R., Tammpere, E., O’Neil, N. J., and Hieter, P. (2020) Cross-species complementation of nonessential yeast genes establishes platforms for testing inhibitors of human proteins, Genetics, 214, 735-747, https://doi.org/10.1534/genetics.119.302971.
9. Qian, Y., Kachroo, A. H., Yellman, C. M., Marcotte, E. M., and Johnson, K. A. (2014) Yeast cells expressing the human mitochondrial DNA polymerase reveal correlations between polymerase fidelity and human disease progression, J. Biol. Chem., 289, 5970-5985, https://doi.org/10.1074/jbc.M113.526418.
10. Parisi, M. A., Xu, B., and Clayton, D. A. (1993) A human mitochondrial transcriptional activator can functionally replace a yeast mitochondrial HMG-box protein both in vivo and in vitro, Mol. Cell Biol., 13, 1951-1961, https://doi.org/10.1128/mcb.13.3.1951-1961.1993.
11. Derbikova, K., Kuzmenko, A., Levitskii, S., Klimontova, M., Chicherin, I., Baleva, M. V., Krasheninnikov, I. A., and Kamenski, P. (2018) Biological and evolutionary significance of terminal extensions of mitochondrial translation initiation factor, Int. J. Mol. Sci., 19, 3861, https://doi.org/10.3390/ijms19123861.
12. Costanzo, M. C., and Fox, T. D. (1988) Specific translational activation by nuclear gene products occurs in the 5' untranslated leader of a yeast mitochondrial mRNA, Proc. Natl. Acad. Sci. USA, 85, 2677-2681, https://doi.org/10.1073/pnas.85.8.2677.
13. Lasserre, J. P., Dautant, A., Aiyar, R. S., Kucharczyk, R., Glatigny, A., Tribouillard-Tanvier, D., Rytka, J., Blondel, M., Skoczen, N., Reynier, P., Pitayu, L., Rotig, A., Delahodde, A., Steinmetz, L. M., Dujardin, G., Procaccio, V., and di Rago, J. P. (2015) Yeast as a system for modeling mitochondrial disease mechanisms and discovering therapies, Dis. Models Mech., 8, 509-526, https://doi.org/10.1242/dmm.020438.
14. Dirick, L., Bendris, W., Loubiere, V., Gostan, T., Gueydon, E., and Schwob, E. (2014) Metabolic and environmental conditions determine nuclear genomic instability in budding yeast lacking mitochondrial DNA, G3 (Bethesda), 4, 411-423, https://doi.org/10.1534/g3.113.010108.
15. Garcia, E. J., Jonge, J. J., Liao, P., Stivison, E., Sing, C. N., Higuchi-Sanabria, R., Boldogh, I. R., and Pon, L. A. (2019) Reciprocal interactions between mtDNA and lifespan control in budding yeast, Mol. Biol. Cell, 30, 2943-2952, https://doi.org/10.1091/mbc.E18-06-0356.
16. Sherman, F. (2002) Getting started with yeast, Methods Enzymol., 350, 3-41, https://doi.org/10.1016/s0076-6879 (02)50954-x.
17. Knop, M., Siegers, K., Pereira, G., Zachariae, W., Winsor, B., Nasmyth, K., and Schiebel, E. (1999) Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines, Yeast, 15, 963-972, https://doi.org/10.1002/ (SICI)1097-0061(199907)15:10B3.0.CO;2-W.
18. Hentges, P., Van Driessche, B., Tafforeau, L., Vandenhaute, J., and Carr, A. M. (2005) Three novel antibiotic marker cassettes for gene disruption and marker switching in Schizosaccharomyces pombe, Yeast, 22, 1013-1019, https://doi.org/10.1002/yea.1291.
19. Gietz, R. D., and Schiestl, R. H. (2007) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method, Nat. Protoc., 2, 31-34, https://doi.org/10.1038/nprot.2007.13.
20. Glick, B. S., and Pon, L. A. (1995) Isolation of highly purified mitochondria from Saccharomyces cerevisiae, Methods Enzymol., 260, 213-223, https://doi.org/10.1016/0076-6879 (95)60139-2.

GOST	Baleva M., Chicherin I., Kamenski P., Khannanov R., Levitskii S., Piunova U., Vasilev R. APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS // Biochemistry. – 2025. – V. 90. – Issue number 9 C. 1325-1337 . URL: https://biochemras.ru/s30345294s0320972525090057-1/?version_id=124145. DOI: 10.7868/S3034529425090057
MLA	Baleva, M.V, Chicherin, I.V, Kamenski, P.A, Khannanov, R.A, Levitskii, S.A, Piunova, U.E, Vasilev, R.A "APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS." Biochemistry. 90.9 (2025).:1325-1337. DOI: 10.7868/S3034529425090057
APA	Baleva M., Chicherin I., Kamenski P., Khannanov R., Levitskii S., Piunova U., Vasilev R. (2025). APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS. Biochemistry. vol. 90, no. 9, pp.1325-1337 DOI: 10.7868/S3034529425090057

RAS BiologyБиохимия Biochemistry

APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS

You can

References

Indexing

RAS BiologyБиохимия Biochemistry

APPROACHES TO HUMANIZATION OF MITOCHONDRIAL PROTEINS IN ON THE EXAMPLE OF REPLACING THE YEAST MITOCHONDRIAL TRANSLATION TERMINATION FACTOR MRF1 WITH ITS HUMAN HOMOLOGS

You can

References

Indexing

Via social network