Rab-Regulated Interaction of Early Endosomes with Lipid Droplets

Pingsheng Liu

René Bartz

John Zehmer

Yun Ying

Meifang Zhu

Ginette Serrero

Richard Anderson

^{Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9039}

^{A&G Pharmaceutical Inc, 9130 Red Branch Rd, Columbia, Maryland 21045}

Address correspondence to: Richard G. W. Anderson Ph.D., Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9039, Tel. 214-648-2346, Fax. 214-648-7577, E-mail: ude.nretsewhtuostu@nosredna.drahcir

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Abstract

Recent studies indicate that lipid droplets isolated from a variety of different cells are rich in proteins known to regulate membrane traffic. Among these proteins are multiple Rab GTPases. Rabs are GTP switches that regulate intracellular membrane traffic through an ability to control membrane-membrane docking as well as vesicle motility. Here we present evidence that the multiple Rabs associated with droplets have a function in regulating membrane traffic. Droplet Rabs are removed by Rab GDP-dissociation inhibitor (RabGDI) in a GDP-dependent reaction, and are recruited to Rab-depleted droplets from cytosol in a GTP-dependent reaction. Rabs also control the recruitment of the early endosome (EE) marker EEA1 from cytosol. We use an in vitro reconstitution assay to show that transferrin receptor positive EEs bind to the droplet in a GTP/Rab-dependent reaction that appears not to lead to membrane fusion. This docking reaction is insensitive to ATP_γs but is blocked by ATP. Finally, we show that when GTP bound active or GDP bound inactive Rab5 is targeted to the droplet, the active form recruits EEA1. We conclude that the Rabs associated with droplets may be capable of regulating the transient interaction of specific membrane systems, probably to transport lipids between membrane compartments.

Abstract

Footnotes

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