Membrane distributions of two ligand-binding receptor complexes in the CLAVATA pathway

Genetic studies have suggested that transmembrane proteins CLAVATA1 (CLV1), CLV2, CORYNE (CRN), BAM1 and BAM2 all play a role in relaying the CLV3 signal and thus regulating stem cell homeostasis at the shoot meristem (SM). The extracellular domain of CLV1 was previously shown to bind the CLE peptide derived from CLV3, providing direct evidence that CLV3-CLV1 function as a ligand-receptor pair. How the other putative receptors function in the CLV pathway, however, remained unclear. We demonstrated in a recent Plant Journal article that the receptor-like protein CLV2 and the receptor-kinases BAM1 and BAM2 also bind to the CLV3 CLE peptide ligand with an affinity similar to that of CLV1. Critically, these ligand binding receptors form two distinct complexes in both transient expression in tobacco and in Arabidopsis meristem cells: a CLV2/CRN multimer and a CLV1/BAM multimer. Here we examine in detail the subcellular membrane partitioning for the receptor proteins in transient expression by two-phase partitioning and co-expression with known subcellular markers. All tested proteins measurably accumulate at the plasma membrane. While CLV1 primarily co-localizes with a plasma membrane marker, CLV2 shows greater co-localization with an endoplasmic reticulum (ER) marker.