Endosidin 2 accelerates PIN2 endocytosis and disturbs intracellular trafficking of PIN2, PIN3, and PIN4 but not of SYT1

Alexandra Lešková; Mária Labajová; Miroslav Krausko; Alexandra Zahradníková; František Baluška; Karol Mičieta; Ján Turňa; Ján Jásik

doi:10.1371/journal.pone.0237448

Endosidin 2 accelerates PIN2 endocytosis and disturbs intracellular trafficking of PIN2, PIN3, and PIN4 but not of SYT1

PLoS One. 2020 Aug 13;15(8):e0237448. doi: 10.1371/journal.pone.0237448. eCollection 2020.

Authors

Alexandra Lešková¹, Mária Labajová¹, Miroslav Krausko¹, Alexandra Zahradníková², František Baluška³, Karol Mičieta^{4

5}, Ján Turňa^{5

6}, Ján Jásik^{1

5}

Affiliations

¹ Institute of Botany, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Bratislava, Slovakia.
² Institute of Experimental Endocrinology, Biomedical Research Center, Slovak Academy of Sciences, Bratislava, Slovakia.
³ Institute of Cellular and Molecular Botany, University of Bonn, Bonn, Germany.
⁴ Department of Botany, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.
⁵ Comenius University Science Park, Comenius University, Bratislava, Slovakia.
⁶ Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.

Abstract

We established that Endosidin2 (ES2) affected the trafficking routes of both newly synthesized and endocytic pools of PIN-FORMED2 (PIN2) in Arabidopsis root epidermal cells. PIN2 populations accumulated in separated patches, which gradually merged into large and compact ES2 aggregates (ES2As). FM4-64 endocytic tracer labeled ES2As as well. Both PIN2 pools also appeared in vacuoles. Accelerated endocytosis of PIN2, its aggregation in the cytoplasm, and redirection of PIN2 flows to vacuoles led to a substantial reduction of the abundance of this protein in the plasma membrane. Whereas PIN-FORMED3 and PIN-FORMED4 also aggregated in the cytoplasm, SYT1 was not sensitive to ES2 treatment and did not appear either in the cytoplasmic aggregates or vacuoles. Ultrastructural analysis revealed that ES2 affects the Golgi apparatus so that stacks acquired cup-shape and even circular shape surrounded by several vesicles. Abnormally shaped Golgi stacks, stack remnants, multi-lamellar structures, separated Golgi cisterna rings, tubular structures, and vesicles formed discrete clusters.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / metabolism*
Arabidopsis Proteins / metabolism*
Cell Wall / metabolism
Cytoplasm / metabolism
Endocytosis / drug effects*
Golgi Apparatus / metabolism
Limonins / pharmacology*
Membrane Transport Proteins / metabolism*
Plant Roots / cytology
Plant Roots / metabolism
Plants, Genetically Modified / metabolism
Protein Transport / drug effects
Synaptotagmin I / metabolism

Substances

Arabidopsis Proteins
Limonins
Membrane Transport Proteins
PIN2 protein, Arabidopsis
PIN3 protein, Arabidopsis
PIN4 protein, Arabidopsis
SYT1 protein, Arabidopsis
Synaptotagmin I
endosidin 2

Grants and funding

This study was supported by Slovak Research and Development Agency (APVV-16-0398 grant awarded to J.J). K.M., J.J and J.T were also supported by the Research and Development Operational Program funded by the ERDF (ITMS 26240220086 grant awarded to J.T).We thank ERFD also for donating the Leica TCS SP8 STED microscope (ITMS 26230120006 grant).The funding bodies were not involved in the design of this study or in the collection of data, analysis, and interpretation of results.