Conformations of the AAA ATPase p97 and its relevance for activity in health and disease

Research Project Summary:

The AAA ATPase p97 is a motor protein converting energy stored in ATP into conformational changes to segregate proteins from large assemblies. Analysis of isolated p97 complexes using single particle cryo-electron microscopy (cryo-SPA) shows that it exists in different conformations. In cells, p97 interacts with up to 20 cofactors that target its function to different pathways. The exact nature, spatial and dynamic properties of co-factors p97 complexes in cell remain unknown. We hypothesize that a better understanding of p97 conformational landscape is critical to understand its functional mechanisms. Therefore, we aim at fully determining and understanding the conformational landscape of isolated p97 complexes, using cryo-SPA and cryo-electron tomography (cryo-ET), and the conformational landscape of p97 complexes inside cells using cryo-ET. We already collaborate on the characterization of the conformational landscape of isolated p97 complexes using cryo-SPA (an ANR-funded project) and, here, we propose to co-supervise 2 PhD students to develop and apply methods to:

  1. Characterize the full conformational landscape of isolated p97 complexes using cryo-ET;
  2. Recognize and characterize the various complexes assembled by p97 inside cells using cryo-ET; and
  3. Combine the conformational landscapes obtained for isolated p97 complexes and in cells to further our understanding of this abundant and essential protein in many cellular processes.

Project Skills and Experience:

The project is interdisciplinary and involves the PI’s expertise in biochemistry and cryo-EM of AAA ATPases (Dr. Rouiller, UoM PI) and cryo-EM image processing methods development (Dr. Jonic, CNRS PI). Also, the PhD students will have complementary backgrounds. The PhD student in the CNRS team will have a background in Computer Science and will be enrolled in the PhD program at the Doctoral School ED 130 – “EDITE de Paris” of Sorbonne Université. The PhD student in the UoM team will have a background in Biochemistry/ Structural Biology and will be enrolled in the PhD program in the Department of Biochemistry and Molecular Biology at the University of Melbourne.

Supervisors:
The University of Melbourne: A/Professor Isabelle Rouiller

Centre national de la recherche scientifique (CNRS): Dr Slavica Jonic

If you are interested in the opportunity, follow the how to apply guidelines.