The cell cycle is tightly controlled by precisely timed and coordinated gene expression. The most profound change in the level of mRNA transcription and polyadenylation can be found in mitosis, in which scanning, and translation elongation are inhibited. In contrast, mTOR signaling remains active, leading to activation of the mRNA cap-binding initiation factor 4E. This unique combination of activating and inhibitory stimuli likely leads to translation of a distinct set of mRNAs.

Our goal is to decipher mitotic role of eIF4E2, which is a member of the 4E cap-binding protein family with both stimulatory and repressive functions in the cell.

The project consists of establishing a stable human RPE cell line with inducible eIF4E2 expression, immunoprecipitation of the labeled eIF4E2 protein, and subsequent LC-MS/MS and Western blot analyses of the interacting eIF4E2 proteins. The second part of the project involves a combination of RNA-seq, bioinformatic and wet-bench approaches to identify and subsequently functionally characterize eIF4E2-bound mRNAs in mitosis.

The candidate should have experience with mammalian cell culture and basic molecular biology techniques. Skills in bioinformatics and fluorescence microscopy/living cell microscopy are highly valued.

The project is fully supported by EU research funds and the Czech Research Grant Agency.

Relevant publications of the research group:

Iyyappan, R., Aleshkina, D., Ming, H., Dvoran, M., Kakavand, K., Jansova, D., Del Llano, E., Gahurova, L., Bruce, A. W., Masek, T., Pospisek, M., Horvat, F., Kubelka, M., Jiang, Z. & Susor, A. The translational oscillation in oocyte and early embryo development. Nucleic Acids Res (2023), doi:10.1093/nar/gkad996.

del Llano, E., Masek, T., Gahurova, L., Pospisek, M., Koncicka, M., Jindrova, A., Jansova, D., Iyyappan, R., Roucova, K., Bruce, A. W., Kubelka, M. & Susor, A. Age-related differences in the translational landscape of mammalian oocytes. Aging Cell 19 (2020), doi:10.1111/acel.13231.

Masek, T., del Llano, E., Gahurova, L., Kubelka, M., Susor, A., Roucova, K., Lin, C.-J., Bruce, A. W. & Pospisek, M. Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method. Int J Mol Sci 21 (2020), doi:10.3390/ijms21041254.

Mrvova, S., Frydryskova, K., Pospisek, M., Vopalensky, V. & Masek, T. Major splice variants and multiple polyadenylation site utilization in mRNAs encoding human translation initiation factors eIF4E1 and eIF4E3 regulate the translational regulators? Molecular Genetics and Genomics 293, 167-186 (2018), doi:10.1007/s00438-017-1375-4.

Frydryskova, K., Masek, T., Borcin, K., Mrvova, S., Venturi, V. & Pospisek, M. Distinct recruitment of human eIF4E isoforms to processing bodies and stress granules. BMC Molecular Biology 17 (2016), doi:10.1186/s12867-016-0072-x.

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