Skip to main content
. 2015 Feb 11;11(2):e1004665. doi: 10.1371/journal.ppat.1004665

Fig 2. ATR1-Cala2 gain of recognition mutations yield RPP1-NdA specific hypersensitive response (HR).

Fig 2

(A) Co-expression of ATR1-Cala2 mutants recovered from random mutagenesis screen with RPP1. Mutant ATR1-expressing Agrobacterium strains were inoculated with RPP1-NdA or RPP1-WsB-expressing strains at OD = 0.45 per construct, with unrecognized (ATR1-Cala2) and recognized (ATR1-Emoy2) alleles as controls. ATR1-Cala2 NdA-GOF comprises E88V/S139T/Y140H/G142R. HR cell death was photographed at 48 hour post inoculation (hpi) for weaker, incomplete response or 72 hpi for a complete response. (B) Western blot of FLAG-tagged ATR1-Cala2, Emoy2, and mutant alleles. Protein was collected at 24 hpi (C) Homology model of ATR1-Cala2 to the seahorse-like ATR1-Emoy2 crystal structure, with “head” and “body” positions of RPP1-NdA specifying mutations (NdA-GOF) in purple and previously identified RPP1-WsB specifying mutations (WsB-GOF) in yellow. An enlarged portion comprising the central “body” WY-motif is shown, with mutations highlighted as above. Y140 was mutated in both NdA- and WsB-GOF ATR1 mutants and is highlighted in yellow. (D) Summary of recognition phenotypes of ATR1 alleles and mutants against both alleles of RPP1.