Fig. 3: TaSGR1 is reduced from an oligomer to a monomer by TaTrx.
From: A fungal pathogen suppresses host leaf senescence to increase infection
a Cysteines at the C-terminal of TaSGR1 are essential for oligomerization. Total proteins extracted from N. benthamiana leaves transiently expressing TaSGR1-GFP, single mutations (C241A, C245A, C247A, C248A, and C253A), and TaSGR15A-GFP (all cysteines replaced by alanine) were detected using the anti-GFP antibody. b Detection of TaSGR1 oligomerization in vitro. TaSGR1-His purified from E. coli and treated with (+) or without (−) 5% β-mercaptoethanol (β-ME) was analyzed via immunoblotting using the anti-His antibody. c Measurement of the disulfide reductase activity of TaTrx using the turbidimetric assay of insulin reduction. Insulin reduction by DTT and GFP served as negative controls. TaTrxM is a mutant where two conserved cysteines in the active site of redox-active disulfide bridges were changed to alanine. Values are mean ± SD, n = 3 biologically independent samples. d The expression of TaTrx in TaSGR1OE#L1 and TaSGR1OE#L3 transgenic wheat lines promoted the transformation of TaSGR1 oligomers to monomers. Total proteins extracted from the leaves of TaSGR1OE#L1 and TaSGR1OE#L3 transgenic wheat plants transiently expressing TaTrx and TaTrxM were treated with or without β-ME and detected using the anti-GFP antibody. e The monomeric TaSGR1 were reduced in Pst CYR31-infected TaSGR1OE plants. Total proteins extracted from the leaves of TaSGR1OE plants were detected using the anti-GFP antibody. Actin proteins detected with the anti-Actin antibody indicated protein loading. Source data are provided as a Source Data file.
