Clathrin-independent endocytosis contributes to uptake of glucose into BY-2 protoplasts

Plant J. 2012 May;70(4):578-84. doi: 10.1111/j.1365-313X.2011.04892.x. Epub 2012 Feb 16.

Abstract

In eukaryotic cells, several pathways exist for the internalization of plasma membrane proteins and extracellular cargo molecules. These endocytic pathways can be divided into clathrin-dependent and clathrin-independent pathways. While clathrin-dependent pathways are known to be involved in a variety of cellular processes in plants, clathrin-independent pathways have so far only been identified in animal and yeast cells. Here we show that internalization of fluorescent glucose into BY-2 cells leads to accumulation of the sugar in compartments of the endocytic pathway. This endocytic uptake of glucose was not blocked by ikarugamycin, an inhibitor of clathrin-dependent endocytosis, suggesting a role for clathrin-independent endocytosis in glucose uptake. Investigations of fusion and fission of single vesicles by membrane capacitance measurements revealed stimulation of endocytic activity by extracellular glucose. Glucose-stimulated fission of vesicles was not affected by addition of ikarugamycin or blocking of clathrin coat formation by transient over-expression of HUB1 (the C-terminal part of the clathrin heavy chain). These data demonstrate that clathrin-independent endocytosis does occur in plant cells. This pathway may represent a common mechanism for the uptake of external nutrients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives
  • 4-Chloro-7-nitrobenzofurazan / metabolism
  • Biological Transport / drug effects
  • Biological Transport / physiology
  • Cell Line
  • Clathrin Heavy Chains / genetics
  • Clathrin Heavy Chains / metabolism
  • Clathrin-Coated Vesicles / metabolism*
  • Cytoplasmic Vesicles / metabolism
  • Cytosol / metabolism
  • Deoxyglucose / analogs & derivatives
  • Deoxyglucose / metabolism
  • Endocytosis / drug effects
  • Endocytosis / physiology*
  • Endosomes / metabolism
  • Glucose / metabolism*
  • Glucose / pharmacology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lactams / pharmacology
  • Microscopy, Confocal
  • Nicotiana / cytology*
  • Nicotiana / metabolism
  • Protoplasts / metabolism*
  • Transfection
  • Vacuoles / metabolism

Substances

  • Lactams
  • Clathrin Heavy Chains
  • Green Fluorescent Proteins
  • ikarugamycin
  • Deoxyglucose
  • 4-Chloro-7-nitrobenzofurazan
  • Glucose
  • 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose