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SHORT COMMUNICATION

The tomato ubiquitin-conjugating enzyme variant Suv, but not SlUev1C and SlUev1D regulates Fen-mediated programmed cell death in Nicotiana benthamiana

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Article: e973814 | Received 19 Jul 2014, Accepted 29 Jul 2014, Published online: 22 Dec 2014

Figures & data

Figure 1. Subcellular localization and bimolecular fluorescence complementation (BiFC) assay using tomato protoplasts. (A). The subcellular localization of Fen, Fni3 and Suv in tomato protoplast. Except for Fen-cGFP in which GFP was fused to the C-terminal of Fen, the GFP protein was in the N terminal in all other fusion proteins. (B) Examination of interaction of Fen and Fni3 with Suv using bimolecular fluorescence complementation (BiFC) assay. Presence of green fluorescence denotes the occurrence of interaction of the 2 proteins in the cell.

Figure 1. Subcellular localization and bimolecular fluorescence complementation (BiFC) assay using tomato protoplasts. (A). The subcellular localization of Fen, Fni3 and Suv in tomato protoplast. Except for Fen-cGFP in which GFP was fused to the C-terminal of Fen, the GFP protein was in the N terminal in all other fusion proteins. (B) Examination of interaction of Fen and Fni3 with Suv using bimolecular fluorescence complementation (BiFC) assay. Presence of green fluorescence denotes the occurrence of interaction of the 2 proteins in the cell.

Figure 2. SlUev1C and SlUev1D are not involved in Fen-mediated PCD in Nicotiana benthamiana. (A) Phylogenetic tree of AtUEV1A-1D and tomato Uev1 homologs. The Amino acid sequences of the proteins were aligned using Clustal W, which was followed by generation of phylogenetic tree using MEGA 6.Citation16,17 (B) Interaction of SlUev1C and SlUev1D with Fni3 and SlUbc13-2, respectively in yeast cells. (C) Alignment of the nucleotide sequence of SlUev1C and 1D using Clustal W. (D) Silencing SlUev1C and SlUev1D gene did not affect Fen-mediated cell death in N. benthamiana leaf.

Figure 2. SlUev1C and SlUev1D are not involved in Fen-mediated PCD in Nicotiana benthamiana. (A) Phylogenetic tree of AtUEV1A-1D and tomato Uev1 homologs. The Amino acid sequences of the proteins were aligned using Clustal W, which was followed by generation of phylogenetic tree using MEGA 6.Citation16,17 (B) Interaction of SlUev1C and SlUev1D with Fni3 and SlUbc13-2, respectively in yeast cells. (C) Alignment of the nucleotide sequence of SlUev1C and 1D using Clustal W. (D) Silencing SlUev1C and SlUev1D gene did not affect Fen-mediated cell death in N. benthamiana leaf.

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