Abstract
The Arabidopsis thaliana genome encodes several genes that are known or predicted to participate in the formation of stress granules (SG). One family of genes encodes for Ras GTPase-activating protein–binding protein (G3BP)-like proteins. Seven genes were identified, of which one of the members was already shown to interact with plant virus proteins in a previous study. A phylogenetic and tissue-specific expression analysis, including laser-dissected phloem, by qRT-PCRs was performed and the sub-cellular localization of individual AtG3BP::EYFP fluorescent fusion proteins expressed in Nicotiana benthamiana epidermal cells was observed. Individual AtG3BP-protein interactions in planta were studied using the bimolecular fluorescence complementation approach in combination with confocal imaging in living cells. In addition, the early and late induction of G3BP-like expression upon Turnip mosaic virus infection was investigated by RNAseq and qRT-PCR. The results showed a high divergence of transcription frequency in the different plant tissues, promiscuous protein–protein interaction within the G3BP-like gene family, and a general induction by a viral infection with TuMV in A. thaliana. The information gained from these studies leads to a better understanding of stress granules, in particular their molecular mode of action in the plant and their role in plant virus infection.
Highlights
The Arabidopsis thaliana genome encodes several genes that are known or predicted to participate in the formation of stress granules (SG)
A protein sequence alignment was performed to show the intra-family relatedness between the postulated members (Fig. 1c). They cluster into two main groups, the first one consisting of AtG3BP-1, -2, -3, and -7, the second one consisting of AtG3BP-4, -5, and -6
AtG3BP-3 amplification was negative in phloem tissue but exclusively expressed in leaf tissue, which might be a hint for guard cell-specific expression
Summary
The Arabidopsis thaliana genome encodes several genes that are known or predicted to participate in the formation of stress granules (SG). The results showed a high divergence of transcription frequency in the different plant tissues, promiscuous protein–protein interaction within the G3BP-like gene family, and a general induction by a viral infection with TuMV in A. thaliana. Abulfaraj and colleagues[26] describe eight family members, generated and analyzed AtG3BP-7 (AT5G48650) OEX lines and KO lines, which showed no phenotype compared to control plants This might be due to the fact that G3BPs in A. thaliana are redundant in their function, and a KO of one AtG3BP could be compensated by one of the others. Aim of this study was to gain more information about all putative members of G3BP-like proteins in A. thaliana, i.e. in terms of tissue expression, cellular localization and protein–protein interaction, and to investigate the response of the AtG3BPs to a viral infection, i.e. TuMV. Special emphasis here was to monitor AtG3BPs expression to an early time point of infection
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