Research Groups

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 ESRF crystallography beamlines are now highly automated and all are equipped with EMBL-designed high-precision diffractometers and frozen crystal sample changers. 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 structural proteomics integrated projects. A very successful robotic system for nanovolume crystallisation has been implemented, and a novel, high-throughput selection method, ESPRIT, has been developed for finding soluble protein fragments from otherwise badly expressed or insoluble proteins. More recently, a Eukaryotic Expression Facility (EEF) has been established to specialise in expression of multi-subunit complexes in insect cells. These platforms are now available to external users under the EU-funded P-CUBE project (www.p-cube.eu). 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. In 2008 a confocal microscope with facilities for cross-correlation spectroscopy was installed for the study of complex formation in cells, and during 2010 a new top-end electron microscope with cryo-tomography capability will become available.

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 protein-DNA interactions and mechanisms of transcriptional regulation is another important topic. Structural analysis of eukaryotic transcription factor DNA 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 elucidate the role of piRNAs in the germ line.

Another major focus is the study of RNA viruses, particularly influenza virus, with the aim of understanding how it replicates and also as a target 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