Molecular interactions of viral proteins with cellular components within and in the vicinity of Potato virus A-induced viroplasm structures

Project Details

Description

The genus Potyvirus contains the largest number of virus species (>150), and is economically the most devastating group of plant viruses. They infect a vast number of plant species, showing that they have evolved very effective strategies for the replication and expression of their genomes. The model virus for this proposed research is Potato virus A (PVA).
Structures called viroplasms provide compartments in which the various stages of the virus multiplication cycle (protein synthesis, RNA replication, virus movement, encapsidation) are likely localized and coordinated. These structures can be regarded as viral factories in the cell. Relatively little is known how the cellular components, like ribosomes and proteasomes, and the different events of virus infection localize within and in the vicinity of these structures and how the transitions from one infection stage to another are coordinated. To that end, the aim of the research proposed is to dissect i) the interactions of viral proteins with the cellular components within and in the vicinity of PVA-induced viroplasm structures and ii) the regulation for transition from viral replication to genome
encapsidation. As viral coat protein (CP) has a central role in regulation of PVA gene expression special attention will be given to investigate the regulation of CP homeostasis and the role of a host chaperones-mediated degradation
pathway during virus infection. Tag-based affinity purification combined with asymmetric flow field fractionation purification for membranous PVA replication complexes and biochemical analysis of the composition of these vesicles combined with confocal and electron tomography imaging of infected cells provides the methodology for studying the composition and contents of the replication-associated structures. When new host components have been identified from the replication complex vesicles, a sensitive luciferase-based quantitation of PVA gene expression combined with overexpression and silencing of host factors will provide essential information about the role of these identified host factors in PVA infection.
StatusFinished
Effective start/end date01/01/201131/12/2014

Funding

  • Unknown funder

Fields of Science

  • 1182 Biochemistry, cell and molecular biology
  • 1183 Plant biology, microbiology, virology
  • Potato virus A
  • Host-virus interactions