Polar auxin transport in the Arabidopsis (Arabidopsis thaliana) root tip maintains high auxin levels around the stem cell niche that gradually decrease in dividing cells but increase again once they transition towards differentiation. Protophloem differentiates earlier than other proximal tissues and employs a unique auxin 'canalization' machinery that is thought to balance auxin efflux with retention. It consists of a proposed activator of PIN-FORMED (PIN) auxin efflux carriers, the AGC kinase PROTEIN KINASE ASSOCIATED WITH BRX (PAX); its inhibitor, BREVIS RADIX (BRX); and PHOSPHATIDYLINOSITOL-4-PHOSPHATE-5-KINASE (PIP5 K) enzymes, which promote polar PAX and BRX localization. Because of dynamic PAX-BRX-PIP5 K interplay, the net cellular output of this machinery remains unclear. Here we deciphered the dosage-sensitive regulatory interactions between PAX, BRX and PIP5 K by their ectopic expression in developing xylem vessels. The data suggest that the dominant collective output of the PAX-BRX-PIP5 K module is a localized reduction in PIN abundance. This requires PAX-stimulated clathrin-mediated PIN endocytosis by site-specific phosphorylation, which distinguishes PAX from other AGC kinases. Ectopic assembly of the PAX-BRX-PIP5 K module is sufficient to cause cellular auxin retention and affects root growth vigor by accelerating the trajectory of xylem vessel development. Our data thus provide direct evidence that local manipulation of auxin efflux alters the timing of cellular differentiation in the root.