Ethylene controls autophosphorylation of the histidine kinase domain in ethylene receptor ETR1.
Perception of the phytohormone ethylene is accomplished by a small family of integral membrane receptors. In Arabidopsis, five ethylene receptor proteins are known, including ethylene resistant 1 (ETR1). The hydrophobic amino-terminal domain of these receptors contains the ethylene-binding site while the carboxyl-terminal part consists of a histidine kinase domain and a response regulator domain, which are well known elements found in bacterial two-component signaling. The soluble membrane-extrinsic carboxyl-terminal part of the receptor, which is likely to play an important role in signal transduction, showed intrinsic kinase activity when expressed and purified on its own. However, a correlation between signal input and autokinase activity was not established in these studies, as receptors were missing the transmembrane amino-terminal sensor domain. Thus, it is still unclear whether autophosphorylation occurs in response to perception of the ethylene signal. Here, we report on autophosphorylation studies of purified full-length ETR1. Autokinase activity of the purified receptor is controlled by ethylene or by ethylene agonists like the pi-acceptor compound cyanide. In fact, both signal molecules were able to completely turn off the intrinsic kinase activity. Furthermore, the observed inhibition of autophosphorylation in ETR1 by both molecules could be prevented when the ethylene antagonist 1-methyl-cyclopropene (MCP) was applied.
Voet-van-Vormizeele
Jan
J
Heinrich-Heine-Universität, Biochemie der Pflanzen, Universitätsstr. 1, 40225 Düsseldorf, Germany.
Groth
Georg
G
D000465:Algorithms; D017360:Arabidopsis / Q000187:drug effects / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002942:Circular Dichroism; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000235:genetics; D005030:Ethylenes / Q000819:agonists / Q000494:pharmacology*; D005822:Genetic Vectors; D000071677:Histidine Kinase; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D017510:Protein Folding / Q000187:drug effects; D011494:Protein Kinases / Q000187:drug effects / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D012966:Sodium Cyanide / Q000494:pharmacology
D029681:Arabidopsis Proteins; D018744:DNA, Plant; C494590:ETR1 protein, Arabidopsis; D005030:Ethylenes; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C036216:ethylene; D011494:Protein Kinases; D000071677:Histidine Kinase; D012966:Sodium Cyanide