Extended Data Fig. 3: Pep1 is a dual function effector and its presence in the Stp complex is essential for virulence.
From: A cell surface-exposed protein complex with an essential virulence function in Ustilago maydis
a, Scheme of Pep1 and Pep1Δ27-42. Violet: signal peptide; black vertical lines: cysteine residues. b, In contrast to the full-length protein, Pep1Δ27-42 does not interact with maize POX12. Serial dilutions of AH109 yeast strains numbered 1 to 6, co-transformed with two plasmids expressing the indicated proteins without signal peptide as fusion to the GAL4 activation domain (AD) or GAL4 binding domain (BD) were grown on SD –Leu –Trp plates for growth control and SD –Leu –Trp –Ade –His plates to assess protein interaction. c, Expression of the fusion proteins in strains 1-6 from (b) was confirmed by HA-specific western blot for AD fusion proteins (top panel) and Myc-specific western blot for BD fusion proteins (lower panel). The open arrow labels AD-HA-Pep1, the closed arrow labels BD-myc-POX12. d, anti-HA western blot of the eluate after HA-specific immunoprecipitation from total extracts of leaves 3 d.p.i. with U. maydis strains expressing the indicated HA-fusion proteins. The arrow marks Pep1. The asterisk marks an unspecific signal. e, Seven-day-old maize seedlings were infected with the indicated U. maydis strains. Disease symptoms were scored at 12 d.p.i. using the disease scores depicted on the bottom. Data represent mean of n = 3 biologically independent experiments. Total numbers of infected plants are indicated above the respective columns.
