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Fig. 2 | BMC Biology

Fig. 2

From: Stability of Begomoviral pathogenicity determinant βC1 is modulated by mutually antagonistic SUMOylation and SIM interactions

Fig. 2

Mutations in βC1 SUMOylation sites abolish pathogenicity. a Design of βC1 SUMOylation motif mutants. SeqLogos indicates conservation in amino acid sequences between βC1. b In vitro SUMOylation assay using purified MBP-βC1 and its SUMOylation motif mutants. Top blot shows SUMOylation conjugates of βC1 probed with anti-SUMO1 and bottom blot is reprobing with anti-MBP. Black arrow indicates intact MBP-βC1. Black and red triangles represent NbSUMO1 and poly-SUMOylated substrates, respectively. c, d Representative phenotypes of GFP-βC1 and its SUMOylation motif mutants in transgenic N. tabacum. c Plants. d Flowers without petals. e Western blot to quantify GFP-βC1 and different SUMOylation motif mutants in transgenic plants using anti-GFP. The star indicates a non-specific band and the arrow shows GFP-βC1. f IP of GFP-βC1 and its SUMOylation motif mutants showing stability of the proteins during transient overexpression. Protein ladder overlaid and false colour applied. g Co-IP of MBP-βC1 and its SUMO mutants during co-overexpression of either NbSUMO1 or NbSUMO1ΔGG. Black and red triangles indicate NbSUMO1 poly-SUMOylated proteins and NbSUMO1-βC1 conjugates, respectively. Protein marker sizes in kDa are indicated. Size bars in c and d are 36 cm (1.2 ft) and 0.8 cm respectively. P, Ponceau staining for total proteins showing RUBISCO

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