Cell Fractionation and the Identification of Host Proteins Involved in Plant–Virus Interactions

• Published in: Pathogens (Basel) Vol. 13; no. 1; p. 53
• Main Authors: Gomaa, Amany E., El Mounadi, Kaoutar, Parperides, Eric, Garcia-Ruiz, Hernan
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.01.2024
• Subjects: Analysis; Antiviral activity; Antiviral agents; antiviral properties; Cell Fractionation; Centrifugation; Chloroplasts; Cloning; Comparative analysis; Control; Crops; Disease resistance; Dosage and administration; Enzymes; Extracellular vesicles; Flowers & plants; Fractionation; genes; Genetic modification; Genome editing; Genomes; Host plants; Host-Pathogen Interactions; Hybridization; Identification and classification; Immunoprecipitation; Infections; Kinases; Localization; Mass Spectrometry; Mass spectroscopy; Mitochondria; Organelles; Pathogenesis; Plant Diseases - virology; Plant Proteins - genetics; Plant Proteins - metabolism; Plant Viruses; precipitin tests; Protein interaction; Proteins; protein–protein interactions; Proteome - metabolism; Proteomes; Proteomics; Proteomics - methods; Replication; RNA polymerase; Roles; subcellular localization; Tobacco; Viral infections; Viral Proteins - genetics; Viral Proteins - metabolism; Virus diseases of plants; Virus Replication; Viruses; virus–host interactions; Yeast; yeasts; United States; cell fractionation; protein–protein interactions; mass spectrometry; subcellular localization; virus–host interactions
• ISSN: 2076-0817; 2076-0817
• DOI: 10.3390/pathogens13010053

Plant viruses depend on host cellular factors for their replication and movement. There are cellular proteins that change their localization and/or expression and have a proviral role or antiviral activity and interact with or target viral proteins. Identification of those proteins and their roles during infection is crucial for understanding plant–virus interactions and to design antiviral resistance in crops. Important host proteins have been identified using approaches such as tag-dependent immunoprecipitation or yeast two hybridization that require cloning individual proteins or the entire virus. However, the number of possible interactions between host and viral proteins is immense. Therefore, an alternative method is needed for proteome-wide identification of host proteins involved in host–virus interactions. Here, we present cell fractionation coupled with mass spectrometry as an option to identify protein–protein interactions between viruses and their hosts. This approach involves separating subcellular organelles using differential and/or gradient centrifugation from virus-free and virus-infected cells (1) followed by comparative analysis of the proteomic profiles obtained for each subcellular organelle via mass spectrometry (2). After biological validation, prospect host proteins with proviral or antiviral roles can be subject to fundamental studies in the context of basic biology to shed light on both virus replication and cellular processes. They can also be targeted via gene editing to develop virus-resistant crops.