Histidine kinase activity of the ETR1 ethylene receptor from Arabidopsis.
ETR1 represents a prototypical ethylene receptor. Homologues of ETR1 have been identified in Arabidopsis as well as in other plant species, indicating that ethylene perception involves a family of receptors and that the mechanism of ethylene perception is conserved in plants. The amino-terminal half of ETR1 contains a hydrophobic domain responsible for ethylene binding and membrane localization. The carboxyl-terminal half of the polypeptide contains domains with homology to histidine kinases and response regulators, signaling motifs originally identified in bacteria. The putative histidine kinase domain of ETR1 was expressed in yeast as a fusion protein with glutathione S-transferase and affinity purified. Autophosphorylation of the purified fusion protein was observed on incubation with radiolabeled ATP. The incorporated phosphate was resistant to treatment with 3 M NaOH, but was sensitive to 1 M HCl, consistent with phosphorylation of histidine. Autophosphorylation was abolished by mutations that eliminated either the presumptive site of phosphorylation (His-353) or putative catalytic residues within the kinase domain. Truncations were used to delineate the region required for histidine kinase activity. An examination of cation requirements indicated that ETR1 requires Mn2+ for autophosphorylation. These results demonstrate that higher plants contain proteins with histidine kinase activity. Furthermore, these results indicate that aspects of ethylene signaling may be regulated by changes in histidine kinase activity of the receptor.
Gamble
R L
RL
Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham, NH 03824, USA.
Coonfield
M L
ML
Schaller
G E
GE
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D005030:Ethylenes / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D000071677:Histidine Kinase; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D010449:Peptide Mapping; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
D005030:Ethylenes; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C083750:ethylene receptors, plant; D008345:Manganese; D006639:Histidine; C036216:ethylene; D011494:Protein Kinases; D000071677:Histidine Kinase; D008274:Magnesium; D002118:Calcium