Endocytosis is an essential cellular process that allows cells to internalise proteins and lipid from the plasma membrane to change its composition and sense and respond to alterations in their extracellular environment. In animal cells, the protein dynamin is involved in membrane scission during endocytosis, allowing invaginating vesicles to become internalised. Arabidopsis encodes two proteins that have all the domains essential for function in the animal dynamins, Dynamin Related Proteins 2A and 2B (DRP2A and 2B). These proteins show very high sequence identity and are both expressed throughout the plant. Single mutants exhibited no obvious phenotypes but double mutants could be recovered as gametophytes carrying mutant copies of both DRP2A and DRP2B were not transmitted to the next generation. Immunolabelling localised DRP2A/B to the tips of root hairs, a site where rapid endocytosis takes place. Constitutive expression of a GTPase defective Dominant Negative form of DRP2A/B did not allow the recovery of plants expressing this protein at a detectable level, demonstrating an interference with endogenous dynamin. Using an inducible expression system Dominant Negative protein was transiently expressed at levels several fold that of the endogenous proteins. Inducible expression of the Dominant Negative protein resulted in reduced endocytosis at the tips of root hairs, as measured by internalisation of an endocytic tracer dye, and resulted in root hairs bulging and bursting. Together these data support a role for DRP2A/B in endocytosis in Arabidopsis, and demonstrates that the function of at least one of these closely related proteins is essential for plant growth.
Plant Molecular Biology – Springer Journals
Published: Apr 3, 2011
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