Extended Data Fig. 5: Conservation of BAK1 C-terminal-tail phosphosites in SERK proteins across plant species.
From: Phosphocode-dependent functional dichotomy of a common co-receptor in plant signalling
a, Clustal Omega multiple alignments were visualized using JalView v2.10.2b2. The alignment is coloured by percentage identity. Yellow, conservation of BAK1 S602, S604 and S612; green, conservation of BAK1 T603. Protein IDs used for the alignments are as follows: Populus trichocarpa PtSERK1(B9MW41), PtSERK2(B9IQM9), PtSERK3(B9HFX1), PtSERK4(B9H599), Dimocarpus longan DlSERK(B5TTV0), Medicago truncatula MtSERK1(Q8GRK2), MtSERK2(E2IXG1), MtSERK3(E2IXG8), MtSERK4(E2IXG2), MtSERK5(E2IXG3), MtSERK6(E2IXG4), Glycine max GmSERK1(C6ZGA8), GmSERK2(C6FF61), Citrus unshiu CuSERK (Q6BE26), Citrus sinensis CsSERK(C3V9W0), Vitis vinifera VvSERK1(D7TXV2), VvSERK2(A5BIY4), VvSERK3(D7STF6), Cyclamen persicum CpSERK1(A7L5U3), CpSERK2(E5D6S9), Gossypium hirsutum GhSERK1(E5Q8K6), GhSERK2(F5BZU9), GhSERK3(F6MF11), Solanum tuberosum StSERK(A3R789), Solanum peruvianum SpSERK(A6N8J2), Solanum lycopersicum SlSERK1(G0XZA3), SlSERK3A(G0XZA5), SlSERK3B(G0XZA6), Daucus carota DcSERK(O23921), Arabidopsis thaliana AtSERK1(Q94AG2), AtSERK2(Q9XIC7), AtSERK3(Q94F62), AtSERK4(Q9SKG5), AtSERK5(Q8LPS5), Nicotiana benthamiana NbSERK3A(E3VXE6), NbSERK3B(E3VXE7), Oryza sativa subsp. Indica OsbiSERK(Q6S7F1), Oryza sativa OsSERK(Q5Y8C8), OsSERKlike1(Q67X31), OsSERKlike2(Q6K4T4), Sorghum bicolor SbSERK1(C5YHV3), SbSERK2(C5Y9S6), SbSERK3(C5XVP5), Ananas comosus AcSERK1(H6SU43), AcSERK2(H6UP78), AcSERK3(H6UP79), Cyrtochilum loxense ClSERK(G2XLB1), Cocos nucifera CnSERK(Q5S1N9), Zea mays ZmSERK1(Q93W70), ZmSERK2(Q94IJ5), ZmSERK3(B4G007), Triticum aestivum TaSERK1(G4XGX1), TaSERK2(G4XGX2), TaSERKlike3(G4XGX3), Selaginella moellendorffii SmSERK1(D8SBB8), SmSERK2(D8S0N3), SmSERK3(D8S4M4), SmSERK4(D8R6C9), Marchantia polymorpha MpSERK(A7VM18), Physcomitrella patens PpSERK1(A9STU8), PpSERK2(A9SMW5), PpSERK3(A9RY79), Poa pratensis PoapSERKlike1(Q659J0), PoapSERKlike2(Q659J1), Closterium ehrenbergii CeSERK(A7VM46). b, WebLogo representation of alignment in a. c, Detection of BAK1 S612 phosphorylation using anti-pS612 specific antibodies on affinity-purified recombinant BAK1 cytoplasmic domain (BAK1CD) following an in vitro kinase assay with cold ATP. Membranes were immunoblotted with anti-pS612 and anti-MBP antibodies. d, BAK1 immunoprecipitation and detection of BAK1 S612 phosphorylation in vivo using anti-pS612 antibodies. Two-week-old seedlings were treated with water (−) or 100 nM flg22 (+) for 10 min. The same membrane was stripped and blotted again with anti-BAK1 antibodies for loading control. e, Western-blot analysis using anti-BAK1 and anti-pS612 antibodies after SDS–PAGE of crude protein extracts from two-week-old seedlings treated with 100 nM flg22 for 10 min. Blots were stained with CBB for loading control. For a–e, all experiments were repeated at least twice with similar results. For blot source data, see Supplementary Fig. 1.
