ppat.1007288.g006.tif (672.63 kB)
Pha13 confers self-ubiquitination E3 ligase and ubiquitin binding ability.
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posted on 2018-09-13, 18:15 authored by 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 YehIn 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.
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dicot Arabidopsis overexpressing orchid Pha 13NPR 1-independent virus defense genesSubcellular localization analysisPha 13immunity Salicylic acidPha 13. Functional domain analysisubiquitin chain binding activityyeast two-hybrid screening1 domainvirus resistanceexogenous SA treatment1 zinc finger domainsE 3 ligaseorchid NPR 1AtSAP 520 domainproteinmaster regulator genePha 13. Pha 13E 3 ligase activity
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