Project: The mystery of mitotic translation
Research group: Pospíšek
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Eukaryotic cell cycle is tightly controlled by the precisely timed mRNA synthesis and activation or degradation of proteins that mediate progression through different cell cycle phases. It has been believed for many years that global translation drops down in mitosis. However, recent experiments using asynchronous cell cultures demonstrate that global translation is only moderately decreased and gene–specific translation occurs instead. In summary, one of the most tempting challenges of the current molecular biology is deciphering molecular mechanisms that govern the mitotic translation initiation and elongation.
The project will include: i) determination of transcript isoforms that are preferentially translated during mitosis. This subproject will employ human cell lines, FACS of asynchronous cell cultures coupled with SSP-profiling (a method developed by us recently) and RNA-seq; ii) proteomic analysis of mitotic translation initiation complex including its post-translational modifications by LC-MS/MS approach; iii) investigation of the cellular role of the eIF4E3 protein, which is a non-canonical member of the cap-binding eIF4E protein family; and iiii) validation of results by in situ hybridization and/or protein localization experiments in order to test the presence of selected mRNAs/proteins in mitotic spindle or its vicinity.
Deadline is closed