bicaudald actively regulates microtubule motor activity in lipid droplet transportbicaudald积极调节微管马达运输活动在脂滴.pdfVIP

BicaudalD Actively Regulates Microtubule Motor Activity in Lipid Droplet Transport Kristoffer S. Larsen, Jing Xu, Silvia Cermelli, Zhanyong Shu, Steven P. Gross* Department of Developmental and Cell Biology, University of California Irvine, Irvine, California, United States of America Abstract Background: A great deal of sub-cellular organelle positioning, and essentially all minus-ended organelle transport, depends on cytoplasmic dynein, but how dynein’s function is regulated is not well understood. BicD is established to play a critical role in mediating dynein function—loss of BicD results in improperly localized nuclei, mRNA particles, and a dispersed Golgi apparatus—however exactly what BicD’s role is remains unknown. Nonetheless, it is widely believed that BicD may act to tether dynein to cargos. Here we use a combination of biophysical and biochemical studies to investigate BicD’s role in lipid droplet transport during Drosophila embryogenesis. Methodology/Principal Findings: Functional loss of BicD impairs the embryo’s ability to control the net direction of droplet transport; the developmentally controlled reversal in transport is eliminated. We find that minimal BicD expression (near- BicDnull) decreases the average run length of both plus and minus end directed microtubule (MT) based transport. A point mutation affecting the BicD N-terminus has very similar effects on transport during cellularization (phase II), but in phase III (gastrulation) motion actually appears better than in the wild-type. Conclusions/Significance: In contrast to a simple static tethering model of BicD function, or a role only in initial dynein recruitment to the cargo, our data uncovers a new dynamic role for BicD in actively regulating transport. Lipid droplets move bi-directionally, and our investigations demonstrate that BicD p