Polarly localized WPR proteins interact with PAN receptors and the actin cytoskeleton during maize stomatal development

Abstract

Polarization of cells prior to asymmetric cell division is crucial for correct cell divisions, cell fate and tissue patterning. In maize stomatal development, polarization of subsidiary mother cells prior to asymmetric division is controlled by the BRK-PAN-ROP pathway. Two catalytically inactive receptor-like kinases, PAN2 and PAN1, are required for correct division plane positioning. Proteins in the BRK-PAN-ROP pathway are polarized in subsidiary mother cells, with the polarization of each protein dependent on the previous one. As most of the known proteins in this pathway do not physically interact, possible interactors that might participate in the pathway are yet to be described. We identified WPR proteins as new players during subsidiary mother cell polarization. WPRs physically interact with PAN receptors, and polarly accumulate in subsidiary mother cells. The polarized localization of WPR proteins depends on PAN2 but not PAN1. CRISPR-Cas9- induced mutations result in division plane defects in subsidiary mother cells, and ectopic expression of WPR-RFP results in stomatal defects and alterations to the actin cytoskeleton. We show certain WPR proteins directly interact with F-actin through their N-terminus. Our data implicate WPR proteins as potentially regulating actin filaments, which providing insight into their molecular function. Together, these results demonstrate that WPR proteins are important for cell polarization. One-sentence summaryFour related proteins, identified via their physical interaction with the receptor PAN2, are polarly localized prior to asymmetric division in stomatal lineage cells, and interact with F-actin.