Development of molecular tools for monitoring and perturbing TORC1 activity in single yeast cells

The Target of Rapamycin (TOR) is a kinase of very high medical relevance, whose structure and function are conserved across all eukaryotes. TOR functions within two functionally and structurally distinct multiprotein complexes, TOR Complex 1 (TORC1) and TOR Complex 2 (TORC2), which are also highly conserved from yeast to humans. In budding yeast, the rapamycin-sensitive TORC1 is the major regulator protein synthesis and the key coordinator of cell growth with the quality and abundance of the available nutrients, especially nitrogen sources.

A major roadblock towards obtaining a clearer view of how yeast TORC1 activity is regulated during the cell cycle is the fact that we are still unable to measure TORC1 activity in single yeast cells. Our goal in this project is to develop and validate a TORC1 biosensor that is suitable for real-time monitoring of TORC1 activity via single-cell microscopy. The developed biosensor will be based on the recently introduced technology of Kinase Translocation Reporters [Regot et al. 2015, Durandau et al. 2015], which convert phosphorylation into nucleocytoplasmic shuttling of a fluorescent protein and thus avoid several technical challenges associated with FRET sensors. The candidate will make use of well-known and characterized yeast TORC1 substrates and nuclear localization sequences to develop several alternative biosensor designs, clone them into yeast and screen them via fluorescence microscopy. The most promising constructs will be characterized in terms of their dynamic properties and will be used to track TORC1 activity during unperturbed growth, as well as upon nutrient shifts.

Candidates should hold a PhD in one of the fields of biochemistry, structural biology or molecular biology. Familiarity with fluorescence microscopy and microfluidics is a plus, and so is experience with cloning in yeast. The candidates should have an excellent command of English (oral and written) and possess excellent communication and collaboration skills.

Fixed-term contract: 24 months.

The University of Groningen offers a salary dependent on qualifications and work experience of € 2,640 gross per month up to a maximum of € 4,166 gross per month (salary scale 10 Dutch Universities) for a full-time job. This position is defined according to the UFO function profile 'researcher 4'. The appointment is temporary for a period of two years.

How to apply
You may apply for this position until 7 October 2018 23:59 h / before 8 October 2018 (Dutch local time) by means of the application form (click on "Apply" below on the advertisement on the university website).

Applications should contain:
• a cv with full publication list
• the full contact details of two references (e.g. thesis supervisors)
• a letter of intent outlining the candidate’s motivation, qualifications, experience and their connection to the above mentioned project.

The preferred starting date will be as soon as possible.

Unsolicited marketing is not appreciated.

Faculty of Science and Engineering

The University of Groningen, located in the north of The Netherlands, enjoys an international reputation as one of the oldest and leading research universities in Europe. The Molecular Systems Biology group (led by Prof Matthias Heinemann), part of the Groningen Biomolecular Sciences and Biotechnology Institute (GBB), hosts the group of Dr Andreas Milias-Argeitis. Dr Milias-Argeitis’ group investigates the role of the TOR kinase during the budding yeast cell cycle. To achieve this, the group uses a combination of computational modeling with state-of-the-art single-cell experimental technologies such as microfluidics and optogenetics, which allow dynamic monitoring and perturbation of single cells under the microscope. The present two-year position will be funded by an NWO Vidi grant recently awarded to Dr Milias-Argeitis.