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Listed: 2024-06-18    
 

Master Thesis  "Engineering the human gamma-TuRC in yeast"

Master thesis at the Center of Molecular Biology and Center of Organismic Studies, Heidelberg University
Groups of Drs. G. Pereira, S. Pfeffer and E. Schiebel
 
The gamma-TuRC is the principal microtubule organizer in human cells.
We resolved its structure using cryo-electron microscopy, revealing how over 40 gamma-TuRC subunits assemble into a 14-spoked helical structure.
In this master's thesis project, we will use budding yeast Saccharomyces cerevisiae as a model system to express all human gamma-TuRC subunits.
First, we will assemble a chromosome derived from synthetic DNA fragments of the gamma-TuRC genes,
in collaboration with the Zeiss Center of Synthetic Genomics at the ZMBH.
Next, we will analyze the assembly of the gamma-TuRC using fluorescence microscopy, biochemical approaches,
single-particle cryo-electron microscopy of purified gamma-TuRC, and cryo-electron tomography of yeast cells.
In the second phase, we will tether the gamma-TuRC to the yeast spindle pole body for functional analysis.
Additionally, we will engineer the gamma-TuRC and study the impact of these structural changes on the microtubule system. 

This project is a collaboration between the laboratories of Drs. G. Pereira, S. Pfeffer, and E. Schiebel.

Please send applications to Dr. Elmar Schiebel (schiebel.elmar@zmbh.uni-heidelberg.de) with a CV, a short letter of motivation, and the names of two referees.

The position is available immediately, and the successful applicant will be supported by a Hiwi contract for six-eight months.

1 Liu P, Zupa E, Neuner A, Bohler A, et al. 2020. Insights into the assembly and activation of the microtubule nucleator gamma-TuRC. Nature 578: 467-71.
2 Zupa E, Zheng A, Neuner A, Würtz M, et al. 2020. The cryo-EM structure of a ?-TuSC elucidates architecture and regulation of minimal microtubule nucleation systems. Nat Commun 11.
3 Böhler A, Vermeulen BJA, Wurtz M, Zupa E, et al. 2021. The gamma-tubulin ring complex: Deciphering the molecular organization and assembly mechanism of a major vertebrate microtubule nucleator. Bioessays.
4 Würtz M, Zupa E, Atorino ES, Annett Neuner A, et al. 2022. Modular assembly of the principal microtubule nucleator ?-TuRC. Nat Comm 473: doi: 10.1038/s41467-022-28079-0.
5 Vermeulen BJ, Bohler A, Gao Q, Neuner A, et al. 2024. gamma-TuRC asymmetry induces local protofilament mismatch at the RanGTP-stimulated microtubule minus end. EMBO J.
6 Hata S, Pastor Peidro A, Panic M, Liu P, et al. 2019. The balance between KIFC3 and EG5 tetrameric kinesins controls the onset of mitotic spindle assembly. Nat Cell Biol 21: 1138-51.
7 Karasu OR, Neuner A, Atorino ES, Pereira G, et al. 2022. The central scaffold protein CEP350 coordinates centriole length, stability, and maturation. J Cell Biol 221

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