a state of the art facility for the study of protein trafficking in vivo
Lead Research Organisation:
University of Leeds
Department Name: Institute of Membrane & Systems Biology
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
Determining protein localisation and dynamics is important for answering many questions in biology. To understand how proteins function and are regulated in vivo, we need approaches by which we can determine where proteins go and when, as well as when and where two proteins interact. New and emerging technologies will go a long way towards helping us answer these questions. The first is the development of photo-activatable GFP (PA-GFP). By tagging proteins with PA-GFP, and then using photo-activation to observe a subset of fluorescently labelled molecules on a low fluorescent background, their fate can be accurately determined. The second is the recent developments in the GFP and RFP fluorophores that have improved behaviour in FRET, enabling us to use this approach to investigate protein-protein interactions in vivo. Furthermore, microscopy is now being developed as a tool for high throughput screening approaches, to investigate the effects of mutations, or for screening large numbers of small molecules for ones that have useful effects in cell biology. The major goal of this application is to upgrade our existing bio-imaging facility into a state-of-the-art facility that can exploit these new technologies, with the focus of studying protein trafficking in vivo.
Publications
Musa H
(2006)
Targeted homozygous deletion of M-band titin in cardiomyocytes prevents sarcomere formation.
in Journal of cell science
Dove B
(2006)
Cell cycle perturbations induced by infection with the coronavirus infectious bronchitis virus and their effect on virus replication.
in Journal of virology
Swailes NT
(2006)
Non-muscle myosins 2A and 2B drive changes in cell morphology that occur as myoblasts align and fuse.
in Journal of cell science
Street M
(2006)
Stimulation of Galphaq-coupled M1 muscarinic receptor causes reversible spectrin redistribution mediated by PLC, PKC and ROCK.
in Journal of cell science
Smith AJ
(2006)
Increased ATP-sensitive K+ channel expression during acute glucose deprivation.
in Biochemical and biophysical research communications
Dalton JE
(2006)
Intraepithelial gammadelta+ lymphocytes maintain the integrity of intestinal epithelial tight junctions in response to infection.
in Gastroenterology
Foresti O
(2006)
Overexpression of the Arabidopsis syntaxin PEP12/SYP21 inhibits transport from the prevacuolar compartment to the lytic vacuole in vivo.
in The Plant cell
Mankouri J
(2006)
Kir6.2 mutations causing neonatal diabetes prevent endocytosis of ATP-sensitive potassium channels.
in The EMBO journal
DaSilva LL
(2006)
Targeting of the plant vacuolar sorting receptor BP80 is dependent on multiple sorting signals in the cytosolic tail.
in The Plant cell
Dove BK
(2006)
Changes in nucleolar morphology and proteins during infection with the coronavirus infectious bronchitis virus.
in Cellular microbiology
Griffiths RA
(2006)
Herpesvirus saimiri-based gene delivery vectors.
in Current gene therapy
Boyne JR
(2006)
Nucleolar trafficking is essential for nuclear export of intronless herpesvirus mRNA.
in Proceedings of the National Academy of Sciences of the United States of America
Boyne JR
(2006)
gamma-2 Herpes virus post-transcriptional gene regulation.
in Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases
Harrison SM
(2007)
Characterisation of cyclin D1 down-regulation in coronavirus infected cells.
in FEBS letters
Reed ML
(2007)
Characterization of the nuclear export signal in the coronavirus infectious bronchitis virus nucleocapsid protein.
in Journal of virology
Xu SZ
(2008)
TRPC channel activation by extracellular thioredoxin.
in Nature
Peckham M
(2008)
Engineering a multi-nucleated myotube, the role of the actin cytoskeleton.
in Journal of microscopy
Mankouri J
(2008)
The hepatitis C virus non-structural protein NS5A alters the trafficking profile of the epidermal growth factor receptor.
in Traffic (Copenhagen, Denmark)
Bubeck J
(2008)
The syntaxins SYP31 and SYP81 control ER-Golgi trafficking in the plant secretory pathway.
in Traffic (Copenhagen, Denmark)
Mankouri J
(2008)
A comparative cell biological analysis reveals only limited functional homology between the NS5A proteins of hepatitis C virus and GB virus B.
in The Journal of general virology
Foresti O
(2008)
Intermediate organelles of the plant secretory pathway: identity and function.
in Traffic (Copenhagen, Denmark)
Dunn S
(2008)
Differential trafficking of Kif5c on tyrosinated and detyrosinated microtubules in live cells.
in Journal of cell science
Xia R
(2008)
Inhibitory interaction between P2X4 and GABA(C) rho1 receptors.
in Biochemical and biophysical research communications
Wilshaw SP
(2008)
Biocompatibility and potential of acellular human amniotic membrane to support the attachment and proliferation of allogeneic cells.
in Tissue engineering. Part A
Description | The aim of this project was to improve our ability to use light microscopy to image cells, and within cells. The funding allowed us to buy additional equipment to upgrade our existing confocal microscopes, so that we could improve our imaging. These microscopes are used by over 20 different research groups within the Faculty of Biological Sciences at the University, and have supported a wide range of research, from imaging organelles and how they move in living plants, to imaging receptors in mammalian cells. |
Exploitation Route | The research can be used by those interested in developing treatments for infections and disease (e.g. pharma companies, clinicians). Imaging is central to understanding the healthy human organism, plants and animals. Without knowledge of how things work, it is very difficult to understand what goes wrong in disease states. The new microscopes are essential for using imaging to understand cellular processes, and in detecting what goes wrong in diseases from virus infections, to inherited mutant |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
URL | http://www.fbs.leeds.ac.uk/facilities/bioimaging/ |
Description | This funding provided an upgrade to our bio-imaging facility which is used by over 40 different research groups across biological sciences and medicine. It has had impact in a broad range of healthcare and biological sciences. |
First Year Of Impact | 2007 |
Sector | Healthcare,Other |
Impact Types | Economic |