%0 Figure %A Chang, Li %A Chang, Ho-Hsiung %A Chang, Jui-Che %A Lu, Hsiang-Chia %A Wang, Tan-Tung %A Hsu, Duen-Wei %A Tzean, Yuh %A Cheng, An-Po %A Chiu, Yi-Shu %A Yeh, Hsin-Hung %D 2018 %T Mapping the ubiquitin-binding region in Pha13 by a yeast two-hybrid assay. %U https://plos.figshare.com/articles/figure/Mapping_the_ubiquitin-binding_region_in_Pha13_by_a_yeast_two-hybrid_assay_/7086608 %R 10.1371/journal.ppat.1007288.g007 %2 https://ndownloader.figshare.com/files/13029971 %K dicot Arabidopsis overexpressing orchid Pha 13 %K NPR 1-independent virus defense genes %K Subcellular localization analysis %K Pha 13 %K immunity Salicylic acid %K Pha 13. Functional domain analysis %K ubiquitin chain binding activity %K yeast two-hybrid screening %K 1 domain %K virus resistance %K exogenous SA treatment %K 1 zinc finger domains %K E 3 ligase %K orchid NPR 1 %K AtSAP 5 %K 20 domain %K protein %K master regulator gene %K Pha 13. Pha 13 %K E 3 ligase activity %X

(A) Left panel indicating Pha13 putative functional domains (A20 and AN1) and the sites of truncation. The ubiquitin-binding region is mapped to the A20 domain of Pha13. Various combinations of Pha13 truncations cloned in pGBK vector were co-transformed with pGADPhaUb into yeast AH109 strain. The transformants were spotted on control plates (2DO: Leucine-Trptophan dropped out medium) and selective plates (3DO/X-Gal: Leucine-Trptophan-Histidine dropped out and X-Gal added medium), which were incubated at 30°C for 4 days before photography. Red stars indicate cysteine in A20 domain. Yellow stars indicate cysteine replaced with lysine. (B) In vitro pull-down assay on AtSAP5 (used as positive control) [44], Pha13 and Pha13 A20 mutant (Pha13A20m), with polyubiquitin chains.

%I PLOS Pathogens