Research
The EMBL Grenoble outstation situated on the EPN campus
The EMBL outstation in Grenoble, France, a laboratory of about 85 people, shares a campus with the European Synchrotron Radiation Facility (ESRF), which produces some of the world’s most intense X-ray beams, and the Institut Laue Langevin (ILL), which provides high-flux neutron beams. The outstation collaborates very closely with these facilities in building and operating beamlines for macromolecular crystallography, in developing the associated instrumentation and techniques and in providing biochemical laboratory facilities and expertise to help external visitors. The highly automated ESRF crystallography beamlines are all equipped with EMBL-designed high-precision microdiractometers and frozen crystal sample changers. One beamline is run by the outstation and the ESRF in collaboration with India. A new X-ray small-angle scattering instrument built by ESRF and EMBL is now operational with a custom designed small-volume automatic sample changer.
High-throughput methods have also been introduced in other steps of the structure determination process, a development closely connected with the outstation’s involvement in several European integrated projects. These include a very successful robotic system for nanovolume crystallisation and a novel, high-throughput screening method, ESPRIT, which enables soluble protein domains to be identified in otherwise badly expressed or insoluble proteins.
The ESRF user facility at the EPN campus
More recently, a Eukaryotic Expression Facility (EEF) has been established specialising in the expression of multi-subunit complexes in insect cells, building on and developing further the well known MultiBac method. These platforms are now available to external users under the EU funded P-CUBE project. They also form part of the Partnership for Structural Biology (PSB), which has been established with the neighbouring ESRF, ILL and the French national Institut de Biologie Structurale (IBS). The PSB is partly housed in a building adjacent to the outstation, together with the CNRS-Grenoble University-EMBL Unit of Virus Host Cell Interactions (UVHCI).
As a result of these local developments, outstation scientists have access to a wide range of techniques including molecular biology and biophysical techniques, cryo-electron microscopy, isotope labelling, NMR, neutron scattering, X-ray crystallography and small angle scattering. A confocal microscope with facilities for cross-correlation spectroscopy is available for the study of complex formation in cells, as well as a new top-end Polara electron microscope with cryo-tomography capability.
A strong tradition at the outstation is the study of systems involving protein-nucleic acid complexes and viruses. The structural work on aminoacyl-tRNA synthetases is particularly well known and has recently focussed on elucidation of the mode of action of a novel boron-containing antibiotic, which targets leucyl-tRNA synthetase. Projects involving protein-RNA interactions also include cryo-EM studies of the signal recognition particle and its interaction with its receptor and the ribosome and other proteins and complexes involved in RNA processing, transport and degradation, such as the nonsense-mediated decay (NMD) pathway. The analysis of mechanisms of transcriptional regulation, including at the epigenetic level, is another important topic. Structural analysis of eukaryotic transcription factor complexes is continuing with groups working on TFIID, complex enhanceosomes and the dosage compensation complex. A molecular cell biology group is also working on the biology of micro-RNAs, in particular trying to understand the biogenesis and role of piRNAs, which are critical for silencing transposons in the germ line.
Another major focus is the study of segmented RNA viruses, particularly influenza and bunya viruses, with the aim of understanding how they replicate and also as targets for anti-viral drug design. Recently the first crystal structures of domains of the influenza virus polymerase have been determined, which depended on the prior identification of soluble fragments using the ESPRIT method.
Stephen Cusack
Head of EMBL Grenoble