RRC ID 6158
Author Fujimoto M, Arimura S, Ueda T, Takanashi H, Hayashi Y, Nakano A, Tsutsumi N.
Title Arabidopsis dynamin-related proteins DRP2B and DRP1A participate together in clathrin-coated vesicle formation during endocytosis.
Journal Proc Natl Acad Sci U S A
Abstract Endocytosis performs a wide range of functions in animals and plants. Clathrin-coated vesicle (CCV) formation is an initial step of endocytosis, and in animal cells is largely achieved by dynamins. However, little is known of its molecular mechanisms in plant cells. To identify dynamin-related proteins (DRPs) involved in endocytic CCV formation in plant cells, we compared the behaviors of two structurally different Arabidopsis DRPs, DRP2B and DRP1A, with those of the clathrin light chain (CLC), a marker of CCVs, at the plasma membrane by variable incidence angle fluorescent microscopy (VIAFM). DRP2B shares domain organization with animal dynamins whereas DRP1A is plant-specific. We show that green fluorescent protein (GFP)-tagged DRP2B and DRP1A colocalized with CLC tagged with monomeric Kusabira Orange (mKO) in Arabidopsis cultured cells. Time-lapse VIAFM observations suggested that both GFP-DRP2B and GFP-DRP1A appeared and accumulated on the existing mKO-CLC foci and disappeared at the same time as or immediately after the disappearance of mKO-CLC. Moreover, DRP2B and DRP1A colocalized and assembled/disassembled together at the plasma membrane in Arabidopsis cells. A yeast two-hybrid assay showed that DRP2B and DRP1A interacted with each other. An inhibitor of clathrin-mediated endocytosis, tyrphostin A23, disturbed the localization of DRP1A, but had little effect on the localization of DRP2B, indicating that DRP1A and DRP2B have different molecular properties. These results suggest that DRP2B and DRP1A participate together in endocytic CCV formation in Arabidopsis cells despite the difference of their molecular properties.
Volume 107(13)
Pages 6094-9
Published 2010-3-30
DOI 10.1073/pnas.0913562107
PII 0913562107
PMID 20231465
PMC PMC2851877
MeSH Arabidopsis / drug effects Arabidopsis / genetics Arabidopsis / metabolism* Arabidopsis Proteins / chemistry Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism* Base Sequence Binding Sites Cell Membrane / metabolism Clathrin Light Chains / metabolism Clathrin-Coated Vesicles / drug effects Clathrin-Coated Vesicles / metabolism* DNA Primers / genetics DNA, Plant / genetics Dynamins / chemistry Dynamins / genetics Dynamins / metabolism* Endocytosis / drug effects Endocytosis / physiology GTP-Binding Proteins / chemistry GTP-Binding Proteins / genetics GTP-Binding Proteins / metabolism* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Microscopy, Fluorescence Models, Biological Molecular Sequence Data Plants, Genetically Modified Protein Structure, Tertiary Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Two-Hybrid System Techniques Tyrphostins / pharmacology
IF 9.58
Times Cited 89
Arabidopsis / Cultured plant cells, genes pda07543