Analysis of interactions among the CLAVATA3 receptors reveals a direct interaction between CLAVATA2 and CORYNE in Arabidopsis.
In Arabidopsis, CORYNE (CRN), a new member of the receptor kinase family, was recently isolated as a key player involved in the CLAVATA3 (CLV3) signaling pathway, thereby playing an important role in regulating the development of shoot and root apical meristems. However, the precise relationships among CLAVATA1 (CLV1), CLAVATA2 (CLV2), and CRN receptors remain unclear. Here, we demonstrate the subcellular localization of CRN and analyze the interactions among CLV1, CLV2, and CRN using firefly luciferase complementation imaging (LCI) assays in both Arabidopsis mesophyll protoplasts and Nicotiana benthamiana leaves. Fluorescence targeting showed that CRN was localized to the plasma membrane. The LCI assays coupled with co-immunoprecipitation assays demonstrated that CLV2 can directly interact with CRN in the absence of CLV3. Additional LCI assays showed that CLV1 did not interact with CLV2, but can interact weakly with CRN. We also found that CLV1 can interact with CLV2-CRN heterodimers, implying that these three proteins may form a complex. Moreover, CRN, rather than CLV1 and CLV2, was able to form homodimers without CLV3 stimulation. Taken together, our results add direct evidence to the newly proposed two-parallel receptor pathways model and therefore provide new insights into the CLV3 signaling pathway.
Zhu
Yingfang
Y
Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100049, China.
Wang
Yuqing
Y
Li
Ruili
R
Song
Xiufen
X
Wang
Qinli
Q
Huang
Shanjin
S
Jin
Jing Bo
JB
Liu
Chun-Ming
CM
Lin
Jinxing
J
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D047468:Immunoprecipitation; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011485:Protein Binding; D055503:Protein Multimerization; D017346:Protein Serine-Threonine Kinases; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface; D015398:Signal Transduction; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014162:Transfection
C118167:AT2G27250 protein, Arabidopsis; D029681:Arabidopsis Proteins; C401621:CLV2 protein, Arabidopsis; D008164:Luminescent Proteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D008156:Luciferases; D020794:Receptor Protein-Tyrosine Kinases; C118166:CLV1 protein, Arabidopsis; C548152:CORYNE protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases