The battle for the 5'end: dissecting a novel virus-specific translation mechanism driven by eIF3
Lead Research Organisation:
The Pirbright Institute
Department Name: Viral Gene Expression
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.
Planned Impact
The preliminary data presented in this application, and the experiments planned to build on our findings, will lead to a step change in our understanding of the regulation of translation by viruses and the host response to infection. This research will have a direct scientific impact in the fields of virology, translational control and virus-host interactions. As mosquito-borne flaviviruses are important human and animal pathogens, our work may identify new targets for treatment of these economically important infections and therefore has the potential to impact on UK health, society and economy.
Industrial and Economic Impact
Understanding the mechanistic details by which microbial pathogens interact with the translation machinery has long been a source of antimicrobial drugs. It has led to the development of highly successful broad-spectrum antibiotics targeting the bacterial ribosome (i.e neomycin, chloramphenicol, tetracycline). Likewise, we will emulate new avenues to develop antivirals that specifically block viral translation for a group of viruses lacking efficient control strategies. Importantly, and beyond viral systems, specialized translation mechanisms relying on canonical factors, carrying out novel regulatory function are increasingly associated with diseases such as cancer. Therefore our work may have broad significance for human health.
In addition, understanding how viruses commandeer the translation machinery has previously enabled for repurposing of viral elements for the design of expression vectors. Gene therapy or protein expression vectors rely on virus-derived IRES to drive gene expression, while polycistronic constructs often include self-cleaving viral protein elements (i.e FMDV 2A). Therefore, identifying a novel mechanism that confers viruses a translational advantage over the host cell will expand the tool set of regulatory elements used in the design of expression or gene therapy vectors.
Public sector and Societal Impact
The flavivirus DENV is the most significant mosquito-borne virus with nearly half the world's population at risk. It is responsible for 390 million infections annually in humans, including 96 million cases of dengue fever and up to 500,000 cases of the potentially fatal haemorrhagic fever. Related flaviviruses are also important pathogens of humans and animals, such as the re-emerged Zika virus associated with developmental disorders or West Nile virus responsible for encephalitis in horses. Our research has the potential to deliver impact by better understanding of these important pathogens of both humans and animals. The findings from our work will be publicised via the University press office and outreach activities to raise awareness in the general public. The co-PI already has good relationships with TV and radio channels through multiple appearances locally, nationally and internationally during coverage of the Zika and Ebola outbreaks.
Training of skilled researchers
Two PDRAs will be recruited and will receive extensive training in modern biochemical and structural techniques to dissect translation. PDRA1 will be trained in molecular biology techniques to study RNA-protein interactions, and virology reverse genetics systems for flaviviruses, including manipulating Schedule 5 pathogens in BSL3 containment facilities. PDRA2 will be equipped the unique skillset of reconstituting translation in vitro from native factors (few laboratories worldwide can do this, including NL/TS in the UK). S/he will also master challenging structural methodologies to dissect biological complexes through collaborating with a leading expert in cryoEM. This holistic set of skills will prepare the PDRAs for challenges relevant to a wide range of careers both in academia or industry, increasing their career prospects. In addition, our laboratories regularly host both undergraduate and post-graduate students, who will also benefit from exposure to the BBSRC funded research.
Industrial and Economic Impact
Understanding the mechanistic details by which microbial pathogens interact with the translation machinery has long been a source of antimicrobial drugs. It has led to the development of highly successful broad-spectrum antibiotics targeting the bacterial ribosome (i.e neomycin, chloramphenicol, tetracycline). Likewise, we will emulate new avenues to develop antivirals that specifically block viral translation for a group of viruses lacking efficient control strategies. Importantly, and beyond viral systems, specialized translation mechanisms relying on canonical factors, carrying out novel regulatory function are increasingly associated with diseases such as cancer. Therefore our work may have broad significance for human health.
In addition, understanding how viruses commandeer the translation machinery has previously enabled for repurposing of viral elements for the design of expression vectors. Gene therapy or protein expression vectors rely on virus-derived IRES to drive gene expression, while polycistronic constructs often include self-cleaving viral protein elements (i.e FMDV 2A). Therefore, identifying a novel mechanism that confers viruses a translational advantage over the host cell will expand the tool set of regulatory elements used in the design of expression or gene therapy vectors.
Public sector and Societal Impact
The flavivirus DENV is the most significant mosquito-borne virus with nearly half the world's population at risk. It is responsible for 390 million infections annually in humans, including 96 million cases of dengue fever and up to 500,000 cases of the potentially fatal haemorrhagic fever. Related flaviviruses are also important pathogens of humans and animals, such as the re-emerged Zika virus associated with developmental disorders or West Nile virus responsible for encephalitis in horses. Our research has the potential to deliver impact by better understanding of these important pathogens of both humans and animals. The findings from our work will be publicised via the University press office and outreach activities to raise awareness in the general public. The co-PI already has good relationships with TV and radio channels through multiple appearances locally, nationally and internationally during coverage of the Zika and Ebola outbreaks.
Training of skilled researchers
Two PDRAs will be recruited and will receive extensive training in modern biochemical and structural techniques to dissect translation. PDRA1 will be trained in molecular biology techniques to study RNA-protein interactions, and virology reverse genetics systems for flaviviruses, including manipulating Schedule 5 pathogens in BSL3 containment facilities. PDRA2 will be equipped the unique skillset of reconstituting translation in vitro from native factors (few laboratories worldwide can do this, including NL/TS in the UK). S/he will also master challenging structural methodologies to dissect biological complexes through collaborating with a leading expert in cryoEM. This holistic set of skills will prepare the PDRAs for challenges relevant to a wide range of careers both in academia or industry, increasing their career prospects. In addition, our laboratories regularly host both undergraduate and post-graduate students, who will also benefit from exposure to the BBSRC funded research.
Organisations
Publications
Ricciardi-Jorge T
(2023)
PKR-mediated stress response enhances dengue and Zika virus replication
in mBio
| Description | We have revealed new details of how flaviviruses control usage of their genomes during the vial replication cycle. We demonstrated that RNA genome conformation changes affect how easily viral protein can be synthesised from the viral genome and that known interactions between the 5' and 3' ends of the genome affect translation efficiency. We also found that binding of the viral RNA-dependent RNA polymerase, NS5, to an RNA structure at the 5' end of the genome can inhibit translation of the genome. Together our findings point to a temporal regulation of genome usage for translation or replication dependent on the levels of specificities viral proteins. |
| Exploitation Route | Our findings reveal new aspects of flavivirus molecular biology and key interactions that occur during infection. As these are specific to the virus it presents novel information that may help guide future therapeutics targeting viral RNA structure and viral protein-RNA interaction. |
| Sectors | Healthcare,Pharmaceuticals and Medical Biotechnology |