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A more recent version of this article appeared on February 1, 2006
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Submitted on May 23, 2005
Revised on October 12, 2005
Accepted on November 6, 2005
Department of Molecular Biology, Princeton University, Princeton, NJ 08544
Monitoring Editor: Trisha Davis
In metazoans, dynein-dependent vesicle transport is mediated by dynactin, containing an actin-related protein, Arp1p, together with a cargo-selection complex containing a second actin-related protein, Arp11. Paradoxically, in budding yeast, models of dynactin function imply an interaction with membranes, while the lack of microtubule-based vesicle transport implied the absence of a cargo-selection complex. Using both genetic and biochemical approaches we demonstrate that Arp10p is the functional yeast homolog of Arp11, suggesting the possible existence of a pointed-end complex in yeast. Specifically, Arp10p interacts with Arp1p and other dynactin subunits and is dependent on Arp1p for stability. Conversely, Arp10p stabilizes the dynactin complex by association with the Arp1p filament pointed-end. Using a novel hRAS-Arp1p one-hybrid assay, we show that Arp1p associates with the plasma membrane dependent on dynactin subunits, but independent of dynein, and sensitive to cell wall damage. We directly show the association of Arp1p with not only the plasma membrane, but also with a less dense membrane fraction. Based on the hRAS-Arp1p assay, loss of Arp10p enhances the apparent association of dynactin with the plasma membrane and suppresses the loss of signaling conferred by cell wall damage.
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