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
Foresti O
(2010)
A recycling-defective vacuolar sorting receptor reveals an intermediate compartment situated between prevacuoles and vacuoles in tobacco.
in The Plant cell
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
Foresti O
(2008)
Intermediate organelles of the plant secretory pathway: identity and function.
in Traffic (Copenhagen, Denmark)
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
Emmott E
(2008)
Viral nucleolar localisation signals determine dynamic trafficking within the nucleolus.
in Virology
Dunn S
(2008)
Differential trafficking of Kif5c on tyrosinated and detyrosinated microtubules in live cells
in Journal of Cell Science
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
Dove B
(2006)
Changes in nucleolar morphology and proteins during infection with the coronavirus infectious bronchitis virus
in Cellular Microbiology
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
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
Dalton JE
(2006)
Intraepithelial gammadelta+ lymphocytes maintain the integrity of intestinal epithelial tight junctions in response to infection.
in Gastroenterology
Bubeck J
(2008)
The syntaxins SYP31 and SYP81 control ER-Golgi trafficking in the plant secretory pathway.
in Traffic (Copenhagen, Denmark)
Brown L
(2011)
A small molecule with differential effects on the PTS1 and PTS2 peroxisome matrix import pathways
in The Plant Journal
Bradley HJ
(2011)
Residues 155 and 348 contribute to the determination of P2X7 receptor function via distinct mechanisms revealed by single-nucleotide polymorphisms.
in The Journal of biological chemistry
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
Boyne JR
(2009)
Nucleolar disruption impairs Kaposi's sarcoma-associated herpesvirus ORF57-mediated nuclear export of intronless viral mRNAs.
in FEBS letters
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
(2010)
Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs.
in The EMBO journal
Bottanelli F
(2011)
Vacuolar transport in tobacco leaf epidermis cells involves a single route for soluble cargo and multiple routes for membrane cargo.
in The Plant cell
Bottanelli F
(2012)
Evidence for sequential action of Rab5 and Rab7 GTPases in prevacuolar organelle partitioning.
in Traffic (Copenhagen, Denmark)
Baker A
(2010)
Peroxisome biogenesis and positioning.
in Biochemical Society transactions
| 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 |