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
Xu SZ
(2008)
TRPC channel activation by extracellular thioredoxin.
in Nature
Xia R
(2008)
Inhibitory interaction between P2X4 and GABA(C) rho1 receptors.
in Biochemical and biophysical research communications
Wu W
(2012)
Different NF-?B activation characteristics of human respiratory syncytial virus subgroups A and B.
in Microbial pathogenesis
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
Whibley C
(2010)
Wild-type and Hupki (human p53 knock-in) murine embryonic fibroblasts: p53/ARF pathway disruption in spontaneous escape from senescence.
in The Journal of biological chemistry
Turrell SJ
(2011)
Mutation of herpesvirus Saimiri ORF51 glycoprotein specifically targets infectivity to hepatocellular carcinoma cell lines.
in Journal of biomedicine & biotechnology
Turrell SJ
(2012)
Cellular uptake of highly-functionalized ruthenium(II) tris-bipyridine protein-surface mimetics.
in Bioorganic & medicinal chemistry letters
Taylor A
(2011)
Mutation of a C-terminal motif affects Kaposi's sarcoma-associated herpesvirus ORF57 RNA binding, nuclear trafficking, and multimerization.
in Journal of virology
Taneja TK
(2009)
Sar1-GTPase-dependent ER exit of KATP channels revealed by a mutation causing congenital hyperinsulinism.
in Human molecular genetics
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 KA
(2011)
Interactions between factor XIII and the alphaC region of fibrinogen.
in Blood
Smith AJ
(2006)
Increased ATP-sensitive K+ channel expression during acute glucose deprivation.
in Biochemical and biophysical research communications
Smith AJ
(2010)
Voltage-dependent charge movement associated with activation of the CLC-5 2Cl-/1H+ exchanger.
in FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Smith AJ
(2008)
Investigation of K(ATP) channel endocytosis by immunofluorescence.
in Methods in molecular biology (Clifton, N.J.)
Smith A
(2010)
Direct endosomal acidification by the outwardly rectifying CLC-5 Cl - /H + exchanger
in The Journal of Physiology
Scott DJ
(2011)
Clot architecture is altered in abdominal aortic aneurysms and correlates with aneurysm size.
in Arteriosclerosis, thrombosis, and vascular biology
Rose K
(2011)
Transcriptional repression of the M channel subunit Kv7.2 in chronic nerve injury.
in Pain
Reed ML
(2007)
Characterization of the nuclear export signal in the coronavirus infectious bronchitis virus nucleocapsid protein.
in Journal of virology
Prandovszky E
(2011)
The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.
in PloS one
Peckham M
(2008)
Engineering a multi-nucleated myotube, the role of the actin cytoskeleton.
in Journal of microscopy
Nyathi Y
(2010)
The Arabidopsis peroxisomal ABC transporter, comatose, complements the Saccharomyces cerevisiae pxa1 pxa2Delta mutant for metabolism of long-chain fatty acids and exhibits fatty acyl-CoA-stimulated ATPase activity.
in The Journal of biological chemistry
Naylor J
(2010)
Pregnenolone sulphate- and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction.
in Circulation research
Musa H
(2006)
Targeted homozygous deletion of M-band titin in cardiomyocytes prevents sarcomere formation.
in Journal of cell science
Milward A
(2010)
Hepatitis C virus NS5A protein interacts with beta-catenin and stimulates its transcriptional activity in a phosphoinositide-3 kinase-dependent fashion.
in The Journal of general virology
McHale R
(2010)
Prussian blue coordination polymer nanobox synthesis using miniemulsion periphery polymerization (MEPP).
in Chemical communications (Cambridge, England)
Manna P
(2010)
Constitutive Endocytic Recycling and Protein Kinase C-mediated Lysosomal Degradation Control KATP Channel Surface Density
in Journal of Biological Chemistry
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)
Mankouri J
(2010)
Enhanced hepatitis C virus genome replication and lipid accumulation mediated by inhibition of AMP-activated protein kinase.
in Proceedings of the National Academy of Sciences of the United States of America
Mankouri J
(2010)
Optineurin negatively regulates the induction of IFNbeta in response to RNA virus infection.
in PLoS pathogens
Mankouri J
(2006)
Kir6.2 mutations causing neonatal diabetes prevent endocytosis of ATP-sensitive potassium channels.
in The EMBO journal
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
Macnab SA
(2011)
Herpesvirus saimiri-mediated delivery of the adenomatous polyposis coli tumour suppressor gene reduces proliferation of colorectal cancer cells.
in International journal of oncology
Li J
(2011)
Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation.
in Circulation research
James NJ
(2010)
The role of Cajal bodies in the expression of late phase adenovirus proteins.
in Virology
Jackson BR
(2011)
An interaction between KSHV ORF57 and UIF provides mRNA-adaptor redundancy in herpesvirus intronless mRNA export.
in PLoS pathogens
Hong Y
(2011)
Herpesvirus saimiri-based endothelin-converting enzyme-1 shRNA expression decreases prostate cancer cell invasion and migration.
in International journal of cancer
Hong Y
(2012)
The impact of amyloid precursor protein signalling and histone deacetylase inhibition on neprilysin expression in human prostate cells.
in International journal of cancer
Hess K
(2012)
A novel mechanism for hypofibrinolysis in diabetes: the role of complement C3.
in Diabetologia
Henderson Z
(2010)
Co-localization of PRiMA with acetylcholinesterase in cholinergic neurons of rat brain: an immunocytochemical study.
in Brain research
Henderson Z
(2010)
Distribution and role of Kv3.1b in neurons in the medial septum diagonal band complex.
in Neuroscience
Harrison SM
(2007)
Characterisation of cyclin D1 down-regulation in coronavirus infected cells.
in FEBS letters
Hall K
(2010)
Unity and diversity in the human adenoviruses: exploiting alternative entry pathways for gene therapy.
in The Biochemical journal
Groppelli E
(2010)
Cell entry of the aphthovirus equine rhinitis A virus is dependent on endosome acidification.
in Journal of virology
Griffiths RA
(2006)
Herpesvirus saimiri-based gene delivery vectors.
in Current gene therapy
Ghosh SR
(2010)
Determination of the mobility of novel and established Caenorhabditis elegans sarcomeric proteins in vivo.
in European journal of cell biology
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 |