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Scientific Fields
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Application Deadline
01
Jun 2022
2022-06-01 8:00:00 2022-06-01 18:00:00 Europe/Paris Postdoctoral Position at the intersection of cell biology and cryo-EM Structural characterization of membrane remodeling proteins and membrane morphologies in vitro and in situ Autophagy is a major recycling pathway that maintains cellular homeostasis by degrading damaged or superfluous cytoplasmic components by a de […] "25-28 rue du Dr. Roux, 75015, Paris" Thomas Wollert thomas.wollert@pasteur.fr

Structural characterization of membrane remodeling proteins and membrane morphologies in vitro and in situ
Autophagy is a major recycling pathway that maintains cellular homeostasis by degrading damaged or superfluous cytoplasmic components by a de novo formed membrane, termed phagophore. Starvation and cytosolic stresses induce non-selective autophagy that recycles bulk cytoplasm instead of specific cargo. The formation of autophagosomes involves massive membrane remodelling and lipid transfer. The molecular foundation and regulation of these processes is currently not well understood. In our most recent study, we identified a protein coat that remodels donor membranes into phagophores (doi.org/10.21203/rs.3.rs-1269772/v1 and Figure 1).

Figure 1: Platinum metal replica electron micrographs of phagophores reconstituted on model membrane in vitro (left) and on cellular membranes in vivo (right). Scale bar = 200 nm (left) and 500 nm (right).

Based on these findings, we offer a Postdoc position to characterize phagophore formation on model membranes in vitro and on cellular membranes in cells using cryo-electron tomography. We established in collaboration with the Nano-Imaging facility of the Institut Pasteur a workflow combining cryo-fluorescent microscopy, focused ion beam milling of cryo-lamellar and cryo-ET to reveal the ultrastructure of membranes and associated proteins in cells and in reconstitutions on model membranes in vitro. The project will involve optimization of the experimental setup, data collection and analysis by sub-tomogram averaging using the established reconstitution protocols for sample preparation.
We offer state of the art facilities including a Vitrobot MK-IV, an Aquilos 2 cryo-FIB, two Glacios Cryo-TEMs, and one Titan Krios cryo-TEM.

We are looking for candidates with experience in cryo-ET and/or data processing by sub-tomogram averaging. Please send your CV with motivation letter and contact details of three referees to: thomas.wollert@pasteur.fr.
Administrative assistent : Sophie Mutricy (sophie.mutricy@pasteur.fr)

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Biochimie Membranaire et Transport
Institut Pasteur
28 rue du Dr Roux
75724 Paris Cedex 15
Téléphone : +33(0) 1 40 61 3146
e-mail : thomas.wollert@pasteur.fr