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The Arabidopsis protein kinase Pto‐interacting 1‐4 is a common target of the oxidative signal‐inducible 1 and mitogen‐activated protein kinases
Author(s) -
Forzani Celine,
Carreri Alessandro,
de la Fuente van Bentem Sergio,
Lecourieux David,
Lecourieux Fatma,
Hirt Heribert
Publication year - 2011
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2011.08033.x
Subject(s) - kinase , immunoprecipitation , protein kinase a , microbiology and biotechnology , arabidopsis , mapk/erk pathway , phosphorylation , serine , mitogen activated protein kinase , mitogen activated protein kinase kinase , protein–protein interaction , arabidopsis thaliana , signal transduction , biochemistry , map kinase kinase kinase , chemistry , biology , gene , mutant
In Arabidopsis thaliana , the serine/threonine protein kinase oxidative signal‐inducible 1 (OXI1), mediates oxidative stress signalling. Its activity is required for full activation of the mitogen‐activated protein kinases (MAPKs), MPK3 and MPK6, in response to oxidative stress. In addition, the serine/threonine protein kinase Pto‐interacting 1‐2 (PTI1‐2) has been positioned downstream from OXI1, but whether PTI1‐2 signals through MAPK cascades is unclear. Using a yeast two‐hybrid screen we show that OXI1 also interacts with PTI1‐4. OXI1 and PTI1‐4 are stress‐responsive genes and are expressed in the same tissues. Therefore, studies were undertaken to determine whether PTI1‐4 is positioned in the OXI1/MAPK signalling pathway. The interaction between OXI1 and PTI1‐4 was confirmed by using in vivo co‐immunoprecipitation experiments. OXI1 and PTI1‐4 were substrates of MPK3 and MPK6 in vitro . Although no direct interaction was detected between OXI1 and MPK3 or MPK6, in vitro binding studies showed interactions between MPK3 or MPK6 with PTI1‐4. In addition, PTI1‐4 and MPK6 were found in vivo in the same protein complex. These results demonstrate that PTI1‐4 signals via OXI1 and MPK6 signalling cascades. Structured digital abstract• PTI1‐4 and OXI1 phosphorylate by protein kinase assay (View interaction)• OXI1 physically interacts with PTI1‐4 by two hybrid (View interaction)• MPK6 physically interacts with PTI1‐4 by anti tag coimmunoprecipitation (View interaction)• MPK3 and OXI1 phosphorylate by protein kinase assay (View interaction)• MPK6 binds to PTI1‐4 by pull down (View interaction)• PTI1‐4 and MPK3 phosphorylate by protein kinase assay (View interaction)• OXI1 phosphorylates OXI1 by protein kinase assay (View interaction)• OXI1 physically interacts with PTI1‐4 by anti tag coimmunoprecipitation (View interaction)• PTI1‐4 and MPK6 phosphorylate by protein kinase assay (View interaction)• PTI1‐4 physically interacts with AGC2‐3 by two hybrid (View interaction)• OXI1 binds to PTI1‐4 by pulldown (Viewinteraction)• MPK6 and OXI1 phosphorylates by protein kinase assay (View interaction)• MPK3 binds to PTI1‐4 by pull down (View interaction)• PTI1‐4 physically interacts with AGC2‐2 by two hybrid (View interaction)• OXI1 physically interacts with PTI1‐1 by two hybrid (View interaction)• PTI1‐4 binds to OXI1 by pull down (View interaction)