Pha13 confers self-ubiquitination E3 ligase and ubiquitin binding ability. Li Chang Ho-Hsiung Chang Jui-Che Chang Hsiang-Chia Lu Tan-Tung Wang Duen-Wei Hsu Yuh Tzean An-Po Cheng Yi-Shu Chiu Hsin-Hung Yeh 10.1371/journal.ppat.1007288.g006 https://plos.figshare.com/articles/figure/Pha13_confers_self-ubiquitination_E3_ligase_and_ubiquitin_binding_ability_/7086605 <p>In vitro ubiquitination analysis was performed on recombinant proteins of Pha13, A20 mutant (Pha13A20m), AN1 mutant (Pha13AN1m), A20/AN1 double mutant (Pha13A20mAN1m) with or without FLAG-tagged ubiquitin (FLAG-Ub), ubiquitin-activating enzyme (E1), or ubiquitin-conjugating enzyme (E2). Ubiquitinated proteins were analyzed with immunoblotting using anti-FLAG antibodies. Coomassie staining was used as a loading control. The ubiquitinated Pha13 (Pha13-2Ub), E1 conjugated with one Ub (E1-Ub), E2 conjugated with one or two Ub (E2-Ub, E2-2Ub), and free Ub are indicated with arrow.</p> 2018-09-13 18:15:16 dicot Arabidopsis overexpressing orchid Pha 13 NPR 1-independent virus defense genes Subcellular localization analysis Pha 13 immunity Salicylic acid Pha 13. Functional domain analysis ubiquitin chain binding activity yeast two-hybrid screening 1 domain virus resistance exogenous SA treatment 1 zinc finger domains E 3 ligase orchid NPR 1 AtSAP 5 20 domain protein master regulator gene Pha 13. Pha 13 E 3 ligase activity