Defining picornaviral replication complexes by molecular virology and state-of-the art imaging - Novel strategies for disease control
Lead Research Organisation:
University of Leeds
Department Name: Sch of Molecular & Cellular Biology
Abstract
Background:
Picornaviruses are responsible for a number of serious diseases, including polio, foot-and-mouth disease (FMDV) and some emerging diseases, e.g. human hand foot and mouth disease. There is an urgent need to develop new therapeutic strategies to address the continuing issue of picornavirus infection. Genome replication, a key therapeutic target, is usually sequestered within 'replication
complexes', thought to improve efficiency and protect this vulnerable part of the lifecycle. However, for picornaviruses, these are poorly defined.
Objectives:
The project is part of a larger research portfolio designed to improve our understanding of important molecular processes at a fundamental level and to develop new picornavirus therapies, supported by research grants from BBSRC, MRC and WHO/Gates foundation and collaborators (UK and overseas).
Novelty:
This project will focus on FMDV, many aspects of the replication of this virus are poorly understood. To our knowledge, no similar work is being undertaken elsewhere.
Timeliness:
We have recently established a reciprocal complementation system to study the cross talk between FMDV genomes [Herod et al, 2015,2016,2017]. These studies have highlighted the importance of multiple co-infection in viral replication. We will exploit this system here, see below.
Experimental approach:
Our work employs replicons - mini-genomes that allow the study of replication in a safe virus-free way which is highly convenient for the study of the mechanics of genome replication. Using reciprocal complementation (together with state-of the art imaging and structural techniques available in-house) we can dissect the functions of both the RNA and protein components of replication complex. i.e.
1. Identifying novel roles of the essential FMDV non-structural proteins and precursors in replication, focusing on the polymerase (3D) and primer of replication (VPg) using mutagenesis and replication inhibitors.
2. Investigating the development and structure of sites of replication (replication factories) in a time-resolved manner e.g. by PALM/STORM and EM.
Picornaviruses are responsible for a number of serious diseases, including polio, foot-and-mouth disease (FMDV) and some emerging diseases, e.g. human hand foot and mouth disease. There is an urgent need to develop new therapeutic strategies to address the continuing issue of picornavirus infection. Genome replication, a key therapeutic target, is usually sequestered within 'replication
complexes', thought to improve efficiency and protect this vulnerable part of the lifecycle. However, for picornaviruses, these are poorly defined.
Objectives:
The project is part of a larger research portfolio designed to improve our understanding of important molecular processes at a fundamental level and to develop new picornavirus therapies, supported by research grants from BBSRC, MRC and WHO/Gates foundation and collaborators (UK and overseas).
Novelty:
This project will focus on FMDV, many aspects of the replication of this virus are poorly understood. To our knowledge, no similar work is being undertaken elsewhere.
Timeliness:
We have recently established a reciprocal complementation system to study the cross talk between FMDV genomes [Herod et al, 2015,2016,2017]. These studies have highlighted the importance of multiple co-infection in viral replication. We will exploit this system here, see below.
Experimental approach:
Our work employs replicons - mini-genomes that allow the study of replication in a safe virus-free way which is highly convenient for the study of the mechanics of genome replication. Using reciprocal complementation (together with state-of the art imaging and structural techniques available in-house) we can dissect the functions of both the RNA and protein components of replication complex. i.e.
1. Identifying novel roles of the essential FMDV non-structural proteins and precursors in replication, focusing on the polymerase (3D) and primer of replication (VPg) using mutagenesis and replication inhibitors.
2. Investigating the development and structure of sites of replication (replication factories) in a time-resolved manner e.g. by PALM/STORM and EM.
