Phosphorylation-Dependent Regulation of G-Protein Cycle during Nodule Formation in Soybean.
Signaling pathways mediated by heterotrimeric G-protein complexes comprising Gα, Gβ, and Gγ subunits and their regulatory RGS (Regulator of G-protein Signaling) protein are conserved in all eukaryotes. We have shown that the specific Gβ and Gγ proteins of a soybean (Glycine max) heterotrimeric G-protein complex are involved in regulation of nodulation. We now demonstrate the role of Nod factor receptor 1 (NFR1)-mediated phosphorylation in regulation of the G-protein cycle during nodulation in soybean. We also show that during nodulation, the G-protein cycle is regulated by the activity of RGS proteins. Lower or higher expression of RGS proteins results in fewer or more nodules, respectively. NFR1 interacts with RGS proteins and phosphorylates them. Analysis of phosphorylated RGS protein identifies specific amino acids that, when phosphorylated, result in significantly higher GTPase accelerating activity. These data point to phosphorylation-based regulation of G-protein signaling during nodule development. We propose that active NFR1 receptors phosphorylate and activate RGS proteins, which help maintain the Gα proteins in their inactive, trimeric conformation, resulting in successful nodule development. Alternatively, RGS proteins might also have a direct role in regulating nodulation because overexpression of their phospho-mimic version leads to partial restoration of nodule formation in nod49 mutants.
Donald Danforth Plant Science Center, St. Louis, Missouri 63132.
Swarup Roy
http://orcid.org/0000-0002-6738-3574
SR
Choudhury
Donald Danforth Plant Science Center, St. Louis, Missouri 63132 spandey@danforthcenter.org.
Sona
http://orcid.org/0000-0002-5570-3120
S
Pandey
D019204:GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D008954:Models, Biological; D010766:Phosphorylation; D010940:Plant Proteins / Q000378:metabolism*; D055170:Plant Root Nodulation / Q000235:genetics; D030821:Plants, Genetically Modified; D011485:Protein Binding / Q000187:drug effects; D020710:RGS Proteins / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D053204:Root Nodules, Plant / Q000235:genetics / Q000254:growth & development*; D013025:Glycine max / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*
D010940:Plant Proteins; D020710:RGS Proteins; D011956:Receptors, Cell Surface; D019204:GTP-Binding Proteins