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© Thomas Wollert

About

Cells of our body need to deal with similar logistical challenges as we do. They need, for instance, transport cargo from one place to another with high fidelity and efficiency within a very short period of time. Moreover, cells need to degrade unwanted or damaged cellular material to prevent accumulation of molecular waste. This, however, imposes another challenge to cells: they need to identify cytoplasmic trash and degrade it in a highly selective manner. All this needs to be achieve in the crowded cytoplasmic environment harbouring a high density of membrane bound organelles such as mitochondria, endoplasmic reticulum, golgi, or lysosomes. The latter are degradative organelles that contain an amanda of lytic enzymes that degrade all material that is delivered to these recycling stations.

How do cells recycle cytoplasmic material?

Cell have evolved a dedicated pathway which selects cargo and transports it to lysosomes for degradation. This pathway has been termed “autophagy” which means self digestion. In autophagy, a crescent shaped membrane is formed which encloses cargo entirely to give rise to an autophagosome that transports its content to lysosomes. Deficits in selective degradation of protein aggregates is closely related to the onset of a number of neurodegenerative diseases such as Alzheimer’s, Parkinson’s or Huntington’s diseases.

Our research therefore aims to reveal molecular mechanisms involved in autophagy, focussing on its relation to the onset of neurodegenerative diseases.To address these questions, we are using a combination of cell biology, biochemistry, high resolution microscopy, electron tomography and genome editing tools.

Upon cytotoxic stress or starvation, however, cells loose specificity and their waste bags start to capture cytoplasm randomly. This process is particularly important during the development of cancer. We could show that a protein shell at the surface of autophagic membranes function as scaffold to support autophagosome maturation. Our hypothesis is that this protein shell functions like a trash bin that physically support the intrinsically dynamic and flexible autophagic membrane.

Autophagy in the test tube

Using in vitro reconstitution techniques is particularly challenging if not only proteins, but also membranes are involved. We established a wide variety of different model membrane systems, including giant, large, and small unilamellar vesicles and supported lipid bilayers, to recapitulate the biogenesis of autophagosomes in the test tube. Fluorescent labeled autophagy related proteins are added to these membranes to reveal their molecular function. We used this technique to reconstitute the autophagic scaffold on membranes and investigated it by fluorescent techniques such as fluorescent recovery after photobleaching.

The movie shows such an experiment with a giant unilamellar vesicles (membrane, red) at which the protein shell (green) was reconstituted. FRAP (fluorescence recovery after photobleaching) revealed that the proteins which make up the shell are immobile, whereas the lipids of the membrane diffuse freely in and out of the bleached area. This example demonstrates the power of our combinatorial approach in discovering fundamental molecular mechanisms in autophagy.

Social Activities

2018:

World Soccer Championships 2018

Football WM 2018 – Quarter Final France – Uruguay

Birthday Cake for Thomas

Thomas’ Birthday

Birthday Cake for Jagan

Jagan’s Birthday

Birthday Cake for Peter

Peter’s Birthday

2017:

Christmas Party

Fête de Noël 2017

Birthday Cake for Stéphane

Stephane’s Birthday

Fundings

Featured publications

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Becoming part of our team

We are a very dynamic, open, interactive, and multidisciplinary research laboratory. We meet the challenges that we are facing in our daily research by using cutting edge techniques including cell culture, fluorescent microscopy, genome editing, recombinant protein expression and purification, reconstitution reactions with model membranes, and structural biology.

Are you worrying that this sounds too much for you?

Don’t worry! Our supportive team members are taking care that you get to handle these techniques. Stèphane, our technician, is very experienced in cell biology and Christine, our engineer, is an expert in protein production. The other team members applying in vitro and in vivo methods to address their scientific questions. Our combinatorial approach is unique as it allows us to address the most difficult and challenging questions in fundamental biolgy.

Become part of our team

We are always looking for new team members who are interested in our research. We encourage highly motivated and talented PhD-students and Postdocs who are experienced in either biophysical / biochemical methods or cell biology to apply. The complementary training in our laboratory will provide you with the opportunity to work at the interface of cell biology, biochemistry and biophysics! Applicants should send a detailed letter explaining their motivation, their previous experience and how they want to contribute to our research along with CV and contact information for three referees to: thomas.wollert@pasteur.fr

Contact

Centre François Jacob, 3er étage, Room 26-03-07b

Email:thomas.wollert@pasteur.fr
Phone: +33 (0) 140 61 31 46
Fax:+33 (0) 140 61 31 48