Identification of important regions for ethylene binding and signaling in the transmembrane domain of the ETR1 ethylene receptor of Arabidopsis.
The ethylene binding domain (EBD) of the Arabidopsis thaliana ETR1 receptor is modeled as three membrane-spanning helices. We surveyed ethylene binding activity in different kingdoms and performed a bioinformatic analysis of the EBD. Ethylene binding is confined to land plants, Chara, and a group of cyanobacteria but is largely absent in other organisms, consistent with our finding that EBD-like sequences are overrepresented among plant and cyanobacterial species. We made amino acid substitutions in 37 partially or completely conserved residues of the EBD and assayed their effects on ethylene binding and signaling. Mutations primarily in residues in Helices I and II midregions eliminated ethylene binding and conferred constitutive signaling, consistent with the inverse-agonist model of ethylene receptor signaling and indicating that these residues define the ethylene binding pocket. The largest class of mutations, clustered near the cytoplasmic ends of Helices I and III, gave normal ethylene binding activity yet still conferred constitutive signaling. Therefore, these residues may play a role in turning off the signal transmitter domain of the receptor. By contrast, only two mutations were loss of function with respect to signaling. These findings yield insight into the structure and function of the EBD and suggest a conserved role of the EBD as a negative regulator of the signal transmitter domain.
Wang
Wuyi
W
Department of Botany, University of Wisconsin, Madison, Wisconsin 53706, USA.
Esch
Jeff J
JJ
Shiu
Shin-Han
SH
Agula
Hasi
H
Binder
Brad M
BM
Chang
Caren
C
Patterson
Sara E
SE
Bleecker
Anthony B
AB
D000595:Amino Acid Sequence; D000596:Amino Acids; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism*; D001419:Bacteria / Q000235:genetics; D005030:Ethylenes / Q000378:metabolism*; D017343:Genes, Plant; D005816:Genetic Complementation Test; D016678:Genome; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism*; D036226:Seedlings / Q000254:growth & development; D016415:Sequence Alignment; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D019076:Transgenes; D015003:Yeasts
D000596:Amino Acids; D029681:Arabidopsis Proteins; C517775:ERS1 protein, Arabidopsis; C494590:ETR1 protein, Arabidopsis; D005030:Ethylenes; D011956:Receptors, Cell Surface; C036216:ethylene