Postdoc SyBOrg, creating a synthetic membrane organelle in E. coli and C reinhardtii
PIs Bruno MIROUX and Francesca ZITO
Postdoc length: 2 years
Work place: Laboratory of Physical and Chemical Biology of Membrane Proteins
Contact: PIs Bruno MIROUX (email@example.com) and Francesca ZITO (firstname.lastname@example.org)
The “revolution” in the resolution of cryo-electron microscopy (cryo-EM) opens new avenues to observe membrane proteins (MPs) in native membranes (1). Our lab contributed in the cryo-electron microscopy analysis of MPs by developing alternative surfactants named amphipol and by the genetic improvement of bacterial expression hosts (2, 3). Here we propose to study membrane biogenesis in the “AtpF model”. The overproduction of the b subunit of the FoF1-ATP synthase, AtpF, induces massive intracellular membrane (ICM) proliferation in C43(DE3) host (4), and preliminary results indicate that the expression of AtpF in the chloroplast of C reinhardtii might also shape its thylakoids membrane organization. Depending of its motivation and competences, the FP-DYNAMO fellow will have the opportunity to develop its project in one of the following aspects: i tomography on both models, Chlamydomonas chloroplast is a perfect material to study the thylakoids architecture by cryo-EM; ii, omic approaches; iii, genetics, by selective disruption of candidate genes involved in membrane biogenesis; iv, biochemical and synthetic biology approaches to set up nanoreactors or nanovesicles for the in vivo cryo-EM analysis of membrane protein complexes (5). The project will benefit from IBPC platforms and from the nanoimaging Cryo-EM facilities at Institut Pasteur and at Padova University.