Subcellular localization of Dnajc14 protein chaperone in the presence and absence of Potato virus A (PVA) infection

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Title: Subcellular localization of Dnajc14 protein chaperone in the presence and absence of Potato virus A (PVA) infection
Author: Borah, Angana
Contributor: University of Helsinki, Faculty of Agriculture and Forestry, Department of Agricultural Sciences
Publisher: Helsingfors universitet
Date: 2017
Language: eng
Thesis level: master's thesis
Discipline: Bioteknik (MAAT)
Biotechnology (MAAT)
Biotekniikka (MAAT)
Abstract: Potato virus A (PVA) is a single stranded RNA virus, belonging to genus Potyvirus. It is commonly found in potato growing countries and it can decrease potato yield up to 40%. PVA replicates in the cytoplasm of the plant cell, but some of its proteins are targeted to the nucleus. Dnajc14 is a protein chaperone found in potato plants and it plays a role in protein folding, complex assembly and molecular disaggregation. These functions might also help RNA virus infection. When nuclear proteome analysis of potato plants was done, it was found that Dnajc14 protein was present in the nucleus of healthy potato cells, while it was absent from the nucleus of PVA infected cells. The objective of this research work was to confirm the subcellular localization of Dnajc14 in healthy and PVA-infected plants. In this research work, Dnajc14 protein was cloned in fusion with Green Fluorescent Protein (Dnajc14-GFP and GFP-Dnajc14) and the localization of the protein was checked in both healthy Nicotiana benthamiana leaves and N. benthamiana leaves systemically infected with PVA using epifluorescence microscope. PVA was tagged with Red Fluorescence Protein in order to identify the infected cells under the microscope. It was observed that in healthy cells, Dnajc14 protein was localized to the nucleus. When various views of the cells in different leaves were checked, it was found that in PVA-infected cells, the number of cells showing nuclear fluorescence was less than in the healthy cells. The decrease in the amounts of Dnajc14 protein in the nucleus of PVA-infected plants was calculated to be about 31%. The results suggest that there is an alteration in the level of this protein chaperone during PVA infection. This change may be beneficial for the viral infection. This report confirms the localization of Dnajc14 in the nucleus of healthy potato plants.
Subject: potato virus A
nuclear cytoplasmic trafficking
Dnajc14 protein chaperone
subcellular localization
epifluorescence microscopy

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