Molecular cloning of ARR genes and characterization of ARR proteins’ E3 ligase interaction domains

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Title: Molecular cloning of ARR genes and characterization of ARR proteins’ E3 ligase interaction domains
Author: Zinchenko, Ganna
Contributor: University of Helsinki, Faculty of Agriculture and Forestry, Department of Agricultural Sciences
Publisher: Helsingfors universitet
Date: 2016
Language: eng
Thesis level: master's thesis
Discipline: Bioteknik (MAAT)
Biotechnology (MAAT)
Biotekniikka (MAAT)
Abstract: The characterization of flower-specific ubiquitin-proteasome system (UPS) components and identification of their functional molecular networks will help to elucidate the involvement of UPS in regulating flower development and/or flowering time and, therefore, reproductive success of the plant. UPS component COP1 ubiquitin E3 ligase is known to regulate flowering time. The earlier data suggests that COP1 could be involved in regulating cytokinin signaling possibly through Arabidopsis Response Regulator1 (ARR1) ubiquitination. ARR1 is a B-type cytokinin response regulator, and it has recently been shown to be an unstable protein. Furthermore, KMD, F-box protein in SCF E3 ligase complex, has been shown to interact with ARR1 as well. The aim of this study is the characterization of COP1 interaction with novel target proteins ARR1, ARR2, ARR10 and ARR12 that appear to be regulated in different ways. Moreover, KMD proteins were included within the study as a possible competitor of COP1 for interaction with ARR1. In order to perform interactome studies, yeast two-hybrid assay with a preceding molecular cloning of the genes of interest was used. The results can be used to unravel the role of ubiquitin mediated regulation of cytokinin pathway.
Subject: Arabidopsis thaliana
Arabidopsis response regulators
cytokinin signaling
ubiquitin proteasome system
yeast two-hybrid

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