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 J
(2010)
Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs
in The EMBO Journal
Naylor J
(2010)
Pregnenolone sulphate- and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction.
in Circulation research
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
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
Mankouri J
(2010)
Optineurin negatively regulates the induction of IFNbeta in response to RNA virus infection.
in PLoS pathogens
Groppelli E
(2010)
Cell entry of the aphthovirus equine rhinitis A virus is dependent on endosome acidification.
in Journal of virology
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
Hall K
(2010)
Unity and diversity in the human adenoviruses: exploiting alternative entry pathways for gene therapy.
in The Biochemical journal
Henderson Z
(2010)
Co-localization of PRiMA with acetylcholinesterase in cholinergic neurons of rat brain: An immunocytochemical study
in Brain Research
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
James NJ
(2010)
The role of Cajal bodies in the expression of late phase adenovirus proteins.
in Virology
Baker A
(2010)
Peroxisome biogenesis and positioning.
in Biochemical Society transactions
Foresti O
(2010)
A recycling-defective vacuolar sorting receptor reveals an intermediate compartment situated between prevacuoles and vacuoles in tobacco.
in The Plant cell
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
Henderson Z
(2010)
Distribution and role of Kv3.1b in neurons in the medial septum diagonal band complex.
in Neuroscience
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)
Direct endosomal acidification by the outwardly rectifying CLC-5 Cl(-)/H(+) exchanger.
in The Journal of physiology
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
Bubeck J
(2008)
The syntaxins SYP31 and SYP81 control ER-Golgi trafficking in the plant secretory pathway.
in Traffic (Copenhagen, Denmark)
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)
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 |