TY - JOUR
T1 - Complete structure of the chemosensory array core signalling unit in an E. coli minicell strain
AU - Burt, Alister
AU - Cassidy, C. Keith
AU - Ames, Peter
AU - Bacia-Verloop, Maria
AU - Baulard, Megghane
AU - Huard, Karine
AU - Luthey-Schulten, Zaida
AU - Desfosses, Ambroise
AU - Stansfeld, Phillip J.
AU - Margolin, William
AU - Parkinson, John S.
AU - Gutsche, Irina
N1 - Funding Information:
This work has received funding from a European Union’s Horizon 2020 research and innovation programme under grant agreement No. 647784 to I.G. Research in the JSP and WM labs was supported by grants GM19559 and GM131705, respectively, from the U.S. National Institute of General Medical Sciences. This work was also supported by the U.S. National Institutes of Health grant P41GM104601 to C.K.C. and Z.L.-S., the U.S. National Science Foundation grant PHY1430124 to C.K.C. and Z.L.-S., and the U.K. Biotechnology and Biological Sciences Research Council grant BB/S003339/1 to C.K.C. and P.J.S. P.J.S.’s lab is also supported by Wellcome (208361/Z/17/Z), the BBSRC (BB/P01948X/1, BB/R002517/1) and the MRC (MR/S009213/1). The Project made use of the ARCHER UK National Supercomputing Service (http://www.archer. ac.uk), provided by HECBioSim, the UK High End Computing Consortium for Biomolecular Simulation (hecbiosim.ac.uk), which is supported by the EPSRC (EP/ L000253/1). We acknowledge Diamond Light Source for access and support of the cryo-EM facilities at the UK’s national Electron Bio-imaging Centre (eBIC), funded by the Wellcome Trust, MRC and BBRSC. Cryo-ET data acquisition has been supported by iNEXT, grant number 653706 (PID:2626 to I.G.), funded by the EU Horizon 2020 programme. For initial minicell characterisation and grid screening, we used the platforms of the Grenoble Instruct-ERIC Center (ISBG: UMS 3518 CNRS-CEA-UGA-EMBL) with support from FRISBI (ANR-10-INSB-05-02) and GRAL, a project of the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003). IBS acknowledges integration into the Interdisciplinary Research Institute of Grenoble (IRIG, CEA). The IBS electron microscope facility is supported by the Rhône-Alpes Region, the Fondation pour la Recherche Médicale (FRM), the fonds FEDER, the Centre National de la Recherche Scientifique (CNRS), the Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA), the University of Grenoble Alpes, EMBL, and the GIS-Infrastructures en Biologie Santé et Agronomie (IBISA). Molecular dynamics simulations were performed on the Blue Waters supercomputer, which is supported by the National Science Foundation (OCI-0725070 and ACI-1238993) and the state of Illinois as part of the Petascale Computational Resource Grant (ACI-1713784). We are particularly grateful to Daniel Clare, Alistair Siebert and Andrew Howe for help with data acquisition at eBIC, and to Daniel Castaño-Diez for help and discussions on image processing and for development of Dynamo. We thank Guy Schoehn for establishing and managing the IBS Grenoble cryo-electron microscopy platform and for providing training and support. We are grateful to Aymeric Peuch for help with the usage of the joint IBS/EMBL Grenoble EM computing cluster.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Motile bacteria sense chemical gradients with transmembrane receptors organised in supramolecular signalling arrays. Understanding stimulus detection and transmission at the molecular level requires precise structural characterisation of the array building block known as a core signalling unit. Here we introduce an Escherichia coli strain that forms small minicells possessing extended and highly ordered chemosensory arrays. We use cryo-electron tomography and subtomogram averaging to provide a three-dimensional map of a complete core signalling unit, with visible densities corresponding to the HAMP and periplasmic domains. This map, combined with previously determined high resolution structures and molecular dynamics simulations, yields a molecular model of the transmembrane core signalling unit and enables spatial localisation of its individual domains. Our work thus offers a solid structural basis for the interpretation of a wide range of existing data and the design of further experiments to elucidate signalling mechanisms within the core signalling unit and larger array.
AB - Motile bacteria sense chemical gradients with transmembrane receptors organised in supramolecular signalling arrays. Understanding stimulus detection and transmission at the molecular level requires precise structural characterisation of the array building block known as a core signalling unit. Here we introduce an Escherichia coli strain that forms small minicells possessing extended and highly ordered chemosensory arrays. We use cryo-electron tomography and subtomogram averaging to provide a three-dimensional map of a complete core signalling unit, with visible densities corresponding to the HAMP and periplasmic domains. This map, combined with previously determined high resolution structures and molecular dynamics simulations, yields a molecular model of the transmembrane core signalling unit and enables spatial localisation of its individual domains. Our work thus offers a solid structural basis for the interpretation of a wide range of existing data and the design of further experiments to elucidate signalling mechanisms within the core signalling unit and larger array.
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U2 - 10.1038/s41467-020-14350-9
DO - 10.1038/s41467-020-14350-9
M3 - Article
C2 - 32029744
AN - SCOPUS:85079056140
SN - 2041-1723
VL - 11
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 743
ER -