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
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
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
Dalton JE
(2006)
Intraepithelial gammadelta+ lymphocytes maintain the integrity of intestinal epithelial tight junctions in response to infection.
in Gastroenterology
Musa H
(2006)
Targeted homozygous deletion of M-band titin in cardiomyocytes prevents sarcomere formation.
in Journal of cell science
Dove B
(2006)
Changes in nucleolar morphology and proteins during infection with the coronavirus infectious bronchitis virus
in Cellular Microbiology
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
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
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
Griffiths RA
(2006)
Herpesvirus saimiri-based gene delivery vectors.
in Current gene therapy
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
Smith AJ
(2006)
Increased ATP-sensitive K+ channel expression during acute glucose deprivation.
in Biochemical and biophysical research communications
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
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
Dunn S
(2008)
Differential trafficking of Kif5c on tyrosinated and detyrosinated microtubules in live cells
in Journal of Cell Science
Xu SZ
(2008)
TRPC channel activation by extracellular thioredoxin.
in Nature
Bubeck J
(2008)
The syntaxins SYP31 and SYP81 control ER-Golgi trafficking in the plant secretory pathway.
in Traffic (Copenhagen, Denmark)
Emmott E
(2008)
Viral nucleolar localisation signals determine dynamic trafficking within the nucleolus.
in Virology
Xia R
(2008)
Inhibitory interaction between P2X4 and GABA(C) rho1 receptors.
in Biochemical and biophysical research communications
Bahnasi YM
(2008)
Modulation of TRPC5 cation channels by halothane, chloroform and propofol.
in British journal of pharmacology
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)
Peckham M
(2008)
Engineering a multi-nucleated myotube, the role of the actin cytoskeleton.
in Journal of microscopy
Smith AJ
(2008)
Investigation of K(ATP) channel endocytosis by immunofluorescence.
in Methods in molecular biology (Clifton, N.J.)
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
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)
Boyne JR
(2009)
Nucleolar disruption impairs Kaposi's sarcoma-associated herpesvirus ORF57-mediated nuclear export of intronless viral mRNAs.
in FEBS letters
Taneja TK
(2009)
Sar1-GTPase-dependent ER exit of KATP channels revealed by a mutation causing congenital hyperinsulinism.
in Human molecular genetics
Dugan GE
(2009)
Dependence of the localization and function of the human cytomegalovirus protein US6 on the transporter associated with antigen processing.
in The Journal of general virology
Emmott E
(2010)
Quantitative proteomics using SILAC coupled to LC-MS/MS reveals changes in the nucleolar proteome in influenza A virus-infected cells.
in Journal of proteome research
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
(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
Foresti O
(2010)
A recycling-defective vacuolar sorting receptor reveals an intermediate compartment situated between prevacuoles and vacuoles in tobacco.
in The Plant cell
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
Mankouri J
(2010)
Optineurin negatively regulates the induction of IFNbeta in response to RNA virus infection.
in PLoS pathogens
Baker A
(2010)
Peroxisome biogenesis and positioning.
in Biochemical Society transactions
Naylor J
(2010)
Pregnenolone sulphate- and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction.
in Circulation research
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
Hall K
(2010)
Unity and diversity in the human adenoviruses: exploiting alternative entry pathways for gene therapy.
in The Biochemical journal
James NJ
(2010)
The role of Cajal bodies in the expression of late phase adenovirus proteins.
in Virology
Boyne JR
(2010)
Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs.
in The EMBO journal
Ghosh SR
(2010)
Determination of the mobility of novel and established Caenorhabditis elegans sarcomeric proteins in vivo.
in European journal of cell biology
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
McHale R
(2010)
Prussian blue coordination polymer nanobox synthesis using miniemulsion periphery polymerization (MEPP).
in Chemical communications (Cambridge, England)
Henderson Z
(2010)
Co-localization of PRiMA with acetylcholinesterase in cholinergic neurons of rat brain: an immunocytochemical study.
in Brain research
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 |