Dr Emmanuel Saridakis

Nationality
Greece
Programme
SMART LOIRE VALLEY GENERAL PROGRAMME
Scientific Field
Period
January, 2017 - January, 2018
Award
LE STUDIUM / Marie Skłodowska-Curie Research Fellowship 

From

Institute of Nanoscience and Nanotechnology N.C.S.R. "DEMOKRITOS" - GR

In residence at

Center for Molecular Biophysics (CBM) / CNRS - FR

Host scientist

Dr Marc Boudvillain

PROJECT

New crystallization strategies for structure-guided pharmacological development – large biological assemblies for rna metabolism

The elucidation of three-dimensional structures of molecular machines that control cellular physiology is necessary for the understanding of the mechanisms of life and for the development of rational screening tests for pharmaceutical applications. Due to the large size of these biological entities and the high resolution which is sought, X-ray crystallographic structure determination is the method of choice. Obtaining crystals of biological complexes however remains often difficult and is the bottleneck to this method. In this project, we will explore new crystallisation strategies and their capacity to be automated with an aim to enhance the likelihood and speed of obtaining crystals of sufficient quality for structure determination. We will apply these strategies to the specially difficult case of complex molecular motors involved in RNA metabolism, some of which are pharmacological targets of great interest. This project has wide application potential for academia (discovery of new biological operating mechanisms), technology (new crystallisation kits) and pharmaceutics (targetted virtual screening).

Events organised by this fellow

Publications in relation with the research project

Publications

Saridakis Emmanuel
Vishwakarma Rishi
Lai-Kee-Him Josephine
Martin Kevin
Simon Isabelle
Cohen-Gonsaud Martin
Coste Franck
Bron Patrick
Margeat Emmanuel
Boudvillain Marc
:
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The bacterial Rho factor is a ring-shaped motor triggering genome-wide transcription termination and R-loop dissociation. Rho is essential in many species, including in Mycobacterium tuberculosis where rho gene inactivation leads to rapid death. Yet, the M. tuberculosis Rho [MtbRho] factor displays poor NTPase and helicase activities, and resistance to the natural Rho inhibitor bicyclomycin [BCM] that remain unexplained. To address these issues, we solved the cryo-EM structure of MtbRho at 3.3 Å resolution. The MtbRho hexamer is poised into a pre-catalytic, open-ring state wherein specific contacts stabilize ATP in intersubunit ATPase pockets, thereby explaining the cofactor preference of MtbRho. We reveal a leucine-to-methionine substitution that creates a steric bulk in BCM binding cavities near the positions of ATP γ-phosphates, and confers resistance to BCM at the expense of motor efficiency. Our work contributes to explain the unusual features of MtbRho and provides a framework for future antibiotic development.