Plant Protein-Mediated Inhibition of Virus Cell-to-Cell Movement: Far-Western Screening and Biological Analysis of a Plant Protein Interacting with a Viral Movement Protein
| Autor: | Nobumitsu Sasaki, Yasuhiko Matsushita, Hiroshi Nyunoya |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine biology fungi food and beverages RNA Plasmodesma biology.organism_classification 01 natural sciences Virus Cell biology Protein–protein interaction 03 medical and health sciences 030104 developmental biology Plant protein Tomato mosaic virus Movement protein Function (biology) 010606 plant biology & botany |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493996346 |
| DOI: | 10.1007/978-1-4939-9635-3_7 |
| Popis: | Cell-to-cell movement via plasmodesmata is a crucial step for plant RNA viruses to determine their host ranges. Many viruses including Tomato mosaic virus (ToMV) encode one or more movement proteins (MPs) that are indispensable for cell-to-cell movement. During movement processes, MPs are thought to interact directly with many plant proteins that may be involved in supporting or inhibiting cell-to-cell movement of viruses. In order to understand the molecular mechanisms that regulate viral spread positively or negatively, it is important to discover such MP-interacting plant proteins and analyze their functions in viral cell-to-cell movement in efficient ways. In this chapter, we provide protocols of a radioisotope-based far-western screening strategy to construct a λ phage cDNA library from a nonhost Brassica campestris (syn. rapa) for ToMV and identify plant proteins that bind directly to the 32P-labeled probe of ToMV MP, and subsequently a biolistic bombardment method to examine whether a plant protein selected have a function as an inhibitory factor that can interfere with virus cell-to-cell movement. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|