Arabidopsis EPSIN1 plays an important role in vacuolar trafficking of soluble cargo proteins in plant cells via interactions with clathrin, AP-1, VTI11, and VSR1.
Epsin and related proteins play important roles in various steps of protein trafficking in animal and yeast cells. Many epsin homologs have been identified in plant cells from analysis of genome sequences. However, their roles have not been elucidated. Here, we investigate the expression, localization, and biological role in protein trafficking of an epsin homolog, Arabidopsis thaliana EPSIN1, which is expressed in most tissues we examined. In the cell, one pool of EPSIN1 is associated with actin filaments, producing a network pattern, and a second pool localizes primarily to the Golgi complex with a minor portion to the prevacuolar compartment, producing a punctate staining pattern. Protein pull-down and coimmunoprecipitation experiments reveal that Arabidopsis EPSIN1 interacts with clathrin, VTI11, gamma-adaptin-related protein (gamma-ADR), and vacuolar sorting receptor1 (VSR1). In addition, EPSIN1 colocalizes with clathrin and VTI11. The epsin1 mutant, which has a T-DNA insertion in EPSIN1, displays a defect in the vacuolar trafficking of sporamin:green fluorescent protein (GFP), but not in the secretion of invertase:GFP into the medium. Stably expressed HA:EPSIN1 complements this trafficking defect. Based on these data, we propose that EPSIN1 plays an important role in the vacuolar trafficking of soluble proteins at the trans-Golgi network via its interaction with gamma-ADR, VTI11, VSR1, and clathrin.
Song
Jinhee
J
Division of Molecular and Life Sciences and Center for Plant Intracellular Trafficking, Pohang University of Science and Technology, Pohang 790-784, Korea.
Lee
Myoung Hui
MH
Lee
Gil-Je
GJ
Yoo
Cheol Min
CM
Hwang
Inhwan
I
D033961:Adaptor Protein Complex 1 / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002966:Clathrin / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D050766:Qb-SNARE Proteins / Q000378:metabolism; D012995:Solubility; D014617:Vacuoles / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000378:metabolism*
D033961:Adaptor Protein Complex 1; D033942:Adaptor Proteins, Vesicular Transport; D029681:Arabidopsis Proteins; D002966:Clathrin; C106564:ELP protein, Arabidopsis; C514753:EPSIN1 protein, Arabidopsis; D050766:Qb-SNARE Proteins; D033921:Vesicular Transport Proteins; C120751:vesicle transport v-SNARE 11 protein, Arabidopsis