Conditional destabilization of the TPLATE complex impairs endocytic internalization

Jie Wang, Klaas Yperman, Peter Grones, Qihang Jiang, Jonathan Dragwidge, Evelien Mylle, Eliana Mor, Jonah Nolf, Dominique Eeckhout, Geert de Jaeger, Bert de Rybel, Roman Pleskot, Daniel van Damme

Research output: Contribution to journalArticleAcademicpeer-review

18 Citations (Scopus)

Abstract

In plants, endocytosis is essential for many developmental and physiological processes, including regulation of growth and development, hormone perception, nutrient uptake, and defense against pathogens. Our toolbox to modulate this process is, however, rather limited. Here, we report a conditional tool to impair endocytosis. We generated a partially functional TPLATE allele by substituting the most conserved domain of the TPLATE subunit of the endocytic TPLATE complex (TPC). This substitution destabilizes TPC and dampens the efficiency of endocytosis. Short-term heat treatment increases TPC destabilization and reversibly delocalizes TPLATE from the plasma membrane to aggregates in the cytoplasm. This blocks FM uptake and causes accumulation of various known endocytic cargoes at the plasma membrane. Short-term heat treatment therefore transforms the partially functional TPLATE allele into an effective conditional tool to impair endocytosis. Next to their role in endocytosis, several TPC subunits are also implicated in actin-regulated autophagosomal degradation. Inactivating TPC via the WDX mutation, however, does not impair autophagy, thus enabling specific and reversible modulation of endocytosis in planta.

Original languageEnglish
Article numbere2023456118
JournalProceedings of the National Academy of Sciences of the United States of America
Volume118
Issue number15
DOIs
Publication statusPublished - 13 Apr 2021
Externally publishedYes

Keywords

  • Arabidopsis
  • Autophagy
  • Clathrin-mediated endocytosis
  • Temperature-sensitive mutant
  • TPLATE complex

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