Activation of AtMPK9 through autophosphorylation that makes it independent of the canonical MAPK cascades

作者：Nagy Szilvia K; Darula Zsuzsanna; Kallai Brigitta M; Boegre Laszlo; Banhegyi Gabor; Medzihradszky Katalin F; Horvath Gabor V; Meszaros Tamas^*

来源：Biochemical Journal, 2015, 467(1): 167-175.

DOI：10.1042/BJ20141176

• AtMPK9
• atypical mitogen-activated protein kinase (MAPK)
• autophosphorylation
• in vitro translation
• mitogen-activated protein kinase kinase (MAPKK)
• plant mitogen-activated protein kinase (MAPK)
• TDY motif

Mitogen-activated protein kinases (MAPKs) are part of conserved signal transduction modules in eukaryotes that are typically organized into three-tiered kinase cascades. The activation of MAPKs in these pathways is fully dependent on the bisphosphorylation of the TXY motif in the T-loop by the pertinent dual-specificity MAPK kinases (MAPKKs). The Arabidopsis mitogen-activated protein kinase 9 (AtMPK9) is a member of an atypical class of MAPKs. Representatives of this MAPK family have a TDY phosphoacceptor site, a long C-terminal extension and lack the common MAPKK-binding docking motif. In the present paper, we describe multiple in vitro and in vivo data showing that AtMPK9 is activated independently of any upstream MAPKKs but rather is activated through autophosphorylation. We mapped the autophosphorylation sites by MS to the TDY motif and to the C-terminal regulatory extension. We mutated the phosphoacceptor sites on the TDY, which confirmed the requirement for bisphorylation at this site for full kinase activity. Next, we demonstrated that the kinase-inactive mutant form of AtMPK9 is not trans-phosphorylated on the TDYsite when mixed with an active AtMPK9, implying that the mechanism of the autocatalytic phosphorylation is intramolecular. Furthermore, we show that in vivo AtMPK9 is activated by salt and is regulated by okadaic acid-sensitive phosphatases. We conclude that the plant AtMPK9 shows similarities to the mammalian atypical MAPKs, such as extracellular-signal-regulated kinase (ERK) 7/8, in terms of an MAPKK-independent activation mechanism.

• 出版日期2015-4-1