Rapid Vacuolar Sorting of the Borate Transporter BOR1 Requires the Adaptor Protein Complex AP-4 in Arabidopsis

Plant Cell Physiol. 2024 Dec 6;65(11):1801-1811. doi: 10.1093/pcp/pcae096.

Abstract

Plants maintain nutrient homeostasis by controlling the activities and abundance of nutrient transporters. In Arabidopsis thaliana, the borate (B) transporter BOR1 plays a role in the efficient translocation of B under low-B conditions. BOR1 undergoes polyubiquitination in the presence of sufficient B and is then transported to the vacuole via multivesicular bodies (MVBs) to prevent B accumulation in tissues at a toxic level. A previous study indicated that BOR1 physically interacts with µ subunits of adaptor protein complexes AP-3 and AP-4, both involved in vacuolar sorting pathways. In this study, we investigated the roles of AP-3 and AP-4 subunits in BOR1 trafficking in Arabidopsis. The lack of AP-3 subunits did not affect either vacuolar sorting or polar localization of BOR1-GFP, whereas the absence of AP-4 subunits resulted in a delay in high-B-induced vacuolar sorting without affecting polar localization. Super-resolution microscopy revealed a rapid sorting of BOR1-GFP into AP-4-positive spots in the trans-Golgi network (TGN) upon high-B supply. These results indicate that AP-4 is involved in sequestration of ubiquitinated BOR1 into a TGN-specific subdomain 'vacuolar-trafficking zone', and is required for efficient sorting of MVB and vacuole. Our findings have thus helped elucidate the rapid vacuolar sorting process facilitated by AP-4 in plant nutrient transporters.

Keywords: Trans-Golgi network; Adaptor protein; BOR1; Membrane trafficking; Protein degradation; Vacuolar sorting.

MeSH terms

  • Adaptor Protein Complex 3 / genetics
  • Adaptor Protein Complex 3 / metabolism
  • Adaptor Protein Complex 4 / genetics
  • Adaptor Protein Complex 4 / metabolism
  • Antiporters* / genetics
  • Antiporters* / metabolism
  • Arabidopsis Proteins* / genetics
  • Arabidopsis Proteins* / metabolism
  • Arabidopsis* / genetics
  • Arabidopsis* / metabolism
  • Boron / metabolism
  • Multivesicular Bodies / metabolism
  • Protein Transport*
  • Ubiquitination
  • Vacuoles* / metabolism
  • trans-Golgi Network* / metabolism

Substances

  • Arabidopsis Proteins
  • BOR1 protein, Arabidopsis
  • Antiporters
  • Adaptor Protein Complex 4
  • Adaptor Protein Complex 3
  • Boron