cDNA sequence of E2 gene family in Arabidopsis thaliana and data of statistical analysis

Liu, Chuanliang1

Published Jan 04, 2024 on Dryad. https://doi.org/10.5061/dryad.tb2rbp07w

Data files

Jan 04, 2024 version files 78.10 KB

Abstract

E2 ubiquitin-conjugating enzymes act as a heart role in the ubiquitination process and are responsible for catalysis ubiquitin transfer. Although the function of ubiquitin-protein ligases (E3s) in plant response to diverse abiotic stress by targeting specific substrates has been well studied, the E2s' involvement in environmental responses and their downstream targets are not well understood. Here, we demonstrated that the E2 ubiquitin-conjugating enzyme 18 (UBC18) regulates the stability of FREE1 to modulate iron deficiency stress. UBC18 affects the ubiquitination of FREE1 and promotes its degradation, overexpression of UBC18 in plants decreases their sensitivity to iron deficiency by reducing the level of FREE1, and high accumulation of FREE1 in the ubc18 mutant resulted in sensitivity to iron deficiency. In addition, we demonstrated the lysine residues K227, K295, K315, and K540 are required for FREE1 ubiquitination and stability regulation, and mutation of these lysines of FREE1 residues resulted in sensitivity to iron starvation in plants. Taken together, our findings reveal a mechanism of UBC18 in response to iron deficiency stress by altering the abundance of FREE1, and further elucidate the role of ubiquitination sites in FREE1 stability regulation and the plant iron deficiency response.

README: cDNA sequence of E2 gene family in Arabidopsis thaliana and data of statistical analysis

https://doi.org/10.5061/dryad.tb2rbp07w

We uploaded the cDNA sequence of E2 gene family in Arabidopsis thaliana in a Word file, and the statistical analysis data used in "UBC18 E2 conjugating enzyme destabilizes the ESCRT component FREE1 in plant iron deficiency responses" are listed in supplementary table S2.

For cDNA sequence of E2 gene family, we use them to generate a phylogenetic tree, which is presented as Fig.S1 in the manuscript.
For the data of Fig 2 in Supplementary Table S2, it means the relative FREE1 protein level indicated in Fig.2F in the manuscript. we first use ImageJ software to analyze the His-Sumo-2HA-FREE1 protein abundance, and the protein level of 0min in Col-0, ubc18-3 and ubc18-2 were set 1 respectively. REP 1# and REP 2# represent two independent experiments.
For the data of Fig 3 in Supplementary Table S2, the data of Fig 3A in Supplementary Table S2 represent the ubiquitination level of FREE1. We use ImageJ software to quantify the abundance of ubiquitinated FREE1 and the GFP-FREE1 protein abundance and analyze the relative percentage of ubiquitinated FREE1 protein.
For the data of Fig 3 in Supplementary Table S2, the data of Fig 3B in Supplementary Table S2 represent the relative protein level of FREE1. We use ImageJ software to quantify the abundance of FREE1 protein and the tubulin protein abundance, and analysis the relative percentage of FREE1 protein in Col-0, ubc18-3 and ubc18-2.
For the data of Fig 3 in Supplementary Table S2, the data of Fig 3D in Supplementary Table S2 represent the relative root length. We use ImageJ software to quantify the root length of Col-0, UBQ:GFP-FREE1, ubc18-3 and ubc18-2 seedlings indicated in Fig.3C in the manuscript, the root length data were quantified by GraphPad Prism software and presented in Fig.3D in the manuscript.
For the data of Fig 6 in Supplementary Table S2, the data of Fig 6C in Supplementary Table S2 represent the relative root length. We use ImageJ software to quantified the root length of Col-0, UBQ:GFP-FREE14G#1, UBQ:GFP-FREE14G#2 indicated in Fig.6C in manuscript, the root length data were quantified by GraphPad Prism software and presented in Fig.6D in manuscript.
For the data of Fig S4 in Supplementary Table S2, the data of Fig S4 in Supplementary Table S2 represent the relative expression of FREE1 gene in Col-0, ubc18-3 plants. the data were quantified by GraphPad Prism software and presented in Fig.S4 in manuscript.
For the data of Fig S5 in Supplementary Table S2, the data of Fig S5 in Supplementary Table S2 represent the relative expression of UBC18 gene in Col-0 plant. the data were quantified by GraphPad Prism software and presented in Fig.S5 in manuscript.