Molecular biology of foot-and-mouth disease virus:entry replication and assembly
Lead Research Organisation:
THE PIRBRIGHT INSTITUTE
Department Name: UNLISTED
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
The proposed research aims to investigate the following key stages of the FMDV life cycle: i) Membrane penetration by using model membranes, ii) Capsid assembly by using fluorescent and structural approaches, iii) Genome replication and packaging by using cell culture & cell-free replication systems. Work will build upon the research findings, hypotheses, collaborations and techniques developed during nine years of picornavirus research at the University of Leeds. This work has involved viruses such as poliovirus (PV), human rhinovirus (HRV) and of particular relevance to this application, the aphthovirus equine rhinitis A virus (ERAV), the latter having been developed specifically as a surrogate model for FMDV. Substantial preliminary data exists for several aspects of this proposal which will expedite rapid progress in i) experimental work, ii) publication in peer reviewed journals and iii) the procurement of external funding with which to expand the program.
Planned Impact
unavailable
Organisations
- THE PIRBRIGHT INSTITUTE (Lead Research Organisation)
- University of Leeds (Collaboration)
- UNIVERSITY OF ST ANDREWS (Collaboration)
- University of Dundee (Collaboration)
- University of Oxford (Collaboration)
- Harvard University (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
- University of Leuven (Collaboration)
- University of Edinburgh (Collaboration)
- Polytechnic University of Valencia (Collaboration)
People |
ORCID iD |
| Tobias Tuthill (Principal Investigator) |
Publications
Fry EE
(2010)
Crystal structure of equine rhinitis A virus in complex with its sialic acid receptor.
in The Journal of general virology
Tuthill TJ
(2010)
Picornaviruses.
in Current topics in microbiology and immunology
Groppelli E
(2010)
Cell entry of the aphthovirus equine rhinitis A virus is dependent on endosome acidification.
in Journal of virology
Walter TS
(2012)
A plate-based high-throughput assay for virus stability and vaccine formulation.
in Journal of virological methods
Tulloch F
(2014)
FMDV replicons encoding green fluorescent protein are replication competent.
in Journal of virological methods
Panjwani A
(2014)
Capsid protein VP4 of human rhinovirus induces membrane permeability by the formation of a size-selective multimeric pore.
in PLoS pathogens
Zhu L
(2016)
Structure of human Aichi virus and implications for receptor binding.
in Nature microbiology
Newman J
(2018)
The Cellular Chaperone Heat Shock Protein 90 Is Required for Foot-and-Mouth Disease Virus Capsid Precursor Processing and Assembly of Capsid Pentamers.
in Journal of virology
Swanson J
(2021)
Generation of Antibodies against Foot-and-Mouth-Disease Virus Capsid Protein VP4 Using Hepatitis B Core VLPs as a Scaffold.
in Life (Basel, Switzerland)
Newman J
(2021)
An Engineered Maturation Cleavage Provides a Recombinant Mimic of Foot-and-Mouth Disease Virus Capsid Assembly-Disassembly.
in Life (Basel, Switzerland)
| Description | Several key findings resulting from this research are summarized here: 1) The first demonstration that the picornavirus capsid protein VP4 induces membrane permeability by the formation of a size-selective multimeric pore. 2) The first demonstration that the picornavirus equine rhinitis A virus (ERAV) is dependent on endosome acidification during the process of cell entry. 3) The development of a novel plate-based high-throughput assay for virus stability and vaccine formulation. 4) Generating and reporting the crystal structure of equine rhinitis a virus in complex with its sialic acid receptor. |
| Exploitation Route | The findings have contributed to further academic research into picornavirus particle alterations, vaccine stability and led to the identification of novel targets such as conserved epitopes suitable for disease control or diagnostics. |
| Sectors | Agriculture Food and Drink Pharmaceuticals and Medical Biotechnology Other |
| Description | The findings have contributed to the development of FMD vaccine stability and led to the identification of novel targets such as conserved epitopes suitable for disease control or diagnostics. |
| First Year Of Impact | 2014 |
| Sector | Agriculture, Food and Drink,Manufacturing, including Industrial Biotechology,Other |
| Impact Types | Societal Economic |
| Description | BBSRC IAA The Pirbright Institute |
| Amount | £300,000 (GBP) |
| Funding ID | BB/S506680/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 03/2018 |
| End | 03/2021 |
| Description | Understanding RNA packaging signals in foot-and-mouth disease virus (FMDV) for improved vaccine production |
| Amount | £428,671 (GBP) |
| Funding ID | BB/V008323/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 04/2021 |
| End | 04/2024 |
| Title | Deep sequencing to identify RNA packaging signals |
| Description | A novel and simple approach to identify predicted RNA secondary structures involved in genome packaging in positive sense RNA viruses (e.g. the picornavirus foot-and-mouth disease virus [FMDV]). By interrogating deep sequencing data generated from both packaged and unpackaged populations of RNA, we have determined multiple regions of the genome with constrained variation in the packaged population. Predicted secondary structures of these regions revealed stem-loops with conservation of structure and a common motif at the loop. Disruption of these features resulted in attenuation of virus growth in cell culture due to a reduction in assembly of mature virions. This study provides evidence for the involvement of predicted RNA structures in picornavirus packaging and offers a readily transferable methodology for identifying packaging requirements in many other viruses. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2017 |
| Provided To Others? | Yes |
| Impact | Impact so far is fundamental knowledge but with several potential avenues for future impact in for example improved vaccines. |
| Description | Dundee |
| Organisation | University of Dundee |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Co-INvestigator on BBSRC sLoLa award |
| Collaborator Contribution | Co-INvestigator on BBSRC sLoLa award |
| Impact | BBSRC sLoLa award ongoing |
| Start Year | 2012 |
| Description | Edinburgh - Haas |
| Organisation | University of Edinburgh |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Co-Investigator on BBSRC sLoLa award Co-Investigator on BBSRC project grant Collaboration in virus-host interactions |
| Collaborator Contribution | Co-Investigator on BBSRC sLoLa award Co-Investigator on BBSRC project grant Collaboration in virus-host interactions |
| Impact | BBSRC sLoLa award BBSRC project grant |
| Start Year | 2012 |
| Description | HMS |
| Organisation | Harvard University |
| Department | Harvard Medical School |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Collaboration |
| Collaborator Contribution | Collaboration |
| Impact | Collaborative research |
| Description | Imperial |
| Organisation | Imperial College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Joint supervision of PhD student |
| Collaborator Contribution | Joint supervision of PhD student |
| Impact | One successful completed PhD student. Papers published or in preparation. Another PhD student in progress. |
| Start Year | 2010 |
| Description | JN - Preparation of capped mesoporpus silica nanoparticles |
| Organisation | Polytechnic University of Valencia |
| Country | Spain |
| Sector | Academic/University |
| PI Contribution | We were involved with testing the nanoparticles once they had been made. We also contributed purified antibody and shipped it to our collaborators for the generation of one type of nanoparticle, which we subsequently tested. We were involved with the intellectual design of experiments |
| Collaborator Contribution | Our partners generated the nanoparticles and shipped them to us for testing. Our partners were also involved with the intellectual design of experiments. |
| Impact | Experimental work performed to investigate the use of mesoporous silica nanoparticles to detect FMDV proteases in clinical samples |
| Start Year | 2016 |
| Description | JS - Production of monoclonal antibodies that recognise FMDV VP4 from mouse spleens |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | VLPs displaying the N-terminal 15 amino acids of FMDV were designed and produced at The Pirbright Institute and then mice were immunised with them. The spleens of the immunised mice were harvested and splenocytes stored. The response to the VP4 sequence was checked by ELISA and the mouse with the best response was selected to send the splenocytes for monoclonal production. |
| Collaborator Contribution | The collaborators took the splenocytes obtained from the mouse experiments and carried out the fusions. They then screened the supernatants for positive wells that detected the VP4 sequence. |
| Impact | Positive wells have been identified indicating some monoclonals that are specific for VP4 have been generated. These will be further screened for ability to recognise virus and to neutralise infection. |
| Start Year | 2017 |
| Description | KU Leuven |
| Organisation | University of Leuven |
| Country | Belgium |
| Sector | Academic/University |
| PI Contribution | collaboration |
| Collaborator Contribution | collaboration |
| Impact | collaboration |
| Start Year | 2012 |
| Description | Leeds FBS |
| Organisation | University of Leeds |
| Department | Faculty of Biological Sciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Research |
| Collaborator Contribution | Research and student supervision |
| Impact | Research |
| Start Year | 2009 |
| Description | Oxford - Strubi |
| Organisation | University of Oxford |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Collaboration on virus structural biology and joint PhD studentship |
| Collaborator Contribution | Collaboration on virus structural biology and joint PhD studentship |
| Impact | Collaboration on virus structural biology has produced a number of novel virus structures and new understanding of virus entry and packaging |
| Start Year | 2008 |
| Description | Roslin - Grey |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Co-investigator in BBSRC project grant Collaboration in genetic screens to understand virus host interactions |
| Collaborator Contribution | Co-investigator in BBSRC project grant Collaboration in genetic screens to understand virus host interactions |
| Impact | Co-investigator in BBSRC project grant Collaboration in genetic screens to understand virus host interactions |
| Start Year | 2019 |
| Description | St Andrews |
| Organisation | University of St Andrews |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Co-investigator on BBSRC sLoLa |
| Collaborator Contribution | Lead-investigator on BBSRC sLoLa |
| Impact | BBSRC sLoLa ongoing |
| Start Year | 2012 |
| Description | Diamond (TT) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | Diamond Light Source Open Day - explaining to general public the importance of structural biology and microscopy for understanding viruses and designing improved vaccines. |
| Year(s) Of Engagement Activity | 2019 |
| Description | GFRA (TT) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Global Foot-and-mouth disease Research Alliance (GFRA) meeting, presentation on virus packaging and implications for vaccine production, initiated collaboration discussions between international academic and industry partners. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Lecture on foot-and-mouth disease virus to Merrist Wood College students |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Undergraduate students |
| Results and Impact | Lecture on foot-and-mouth disease virus to Merrist Wood College students |
| Year(s) Of Engagement Activity | 2013 |
| Description | SinoPic TT |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | 'SinoPic': Structural Biology of Picornaviruses meeting in China. Generating interest and collaborations in virus structural biology within China. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Surrey Science And Technology Regional Organisation event at Brooklands |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Hopefully inspired children to think about science |
| Year(s) Of Engagement Activity | 2013 |