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. 2018 Jun 5;30(7):1543–1561. doi: 10.1105/tpc.17.00981

Figure 1.

Figure 1.

Patterns Trigger MAPKKK3/5 Phosphorylation.

(A) Schematic representation of MAPKKK5 domain structure and deletion constructs. Green, N-terminal domain (N); orange, kinase domain (KD); aqua, C-terminal tail (T). The C terminus construct (C) contains both the KD and T.

(B) Flg22-induced phosphorylation of MAPKKK5 in the C terminus. Protoplasts expressing full-length, truncated, or kinase-dead (K375M) variants of MAPKKK5 were treated with flg22. Total protein treated with (+) or without (−) λ protein phosphatase (PPase) was subjected to SDS-PAGE containing Phos-tag acrylamide and detected by immunoblotting with anti-HA antibody. Ponceau staining of Rubisco indicates equal loading.

(C) and (D) Pattern-triggered phosphorylation of MAPKKK3 and MAPKKK5 C termini. Protoplasts expressing C-MAPKKK5K375M-HA (C) and C-MAPKKK3K243M-HA (D) were treated with chitin, elf18, Pep2, or flg22. Total protein was treated with (+) or without (−) λ protein phosphatase (PPase) and subjected to SDS-PAGE containing Phos-tag acrylamide and detected by immunoblotting with anti-HA antibody.