TY - JOUR
T1 - Dynactin-dependent, dynein-driven vesicle transport in the absence of membrane proteins
T2 - A role for spectrin and acidic phospholipids
AU - Muresan, Virgil
AU - Stankewich, Michael C.
AU - Steffen, Walter
AU - Morrow, Jon S.
AU - Holzbaur, Erika L.F.
AU - Schnapp, Bruce J.
N1 - Funding Information:
We are grateful to Dr. Lewis Cantley, Dr. Tom Rapoport, and Dr. Thomas S. Reese for their continual support and advice. We also thank Asya Lyass for technical assistance and Nathaniel Allen, Dr. Judy Chou, Corinne John, Dr. Song Jae Kil, Martin Köhrmann, Dr. Sunjong Kwon, Dr. Trent Munro, and Dr. Kristen Verhey for comments on the manuscript. We are grateful to Dr. Thomas S. Reese (Laboratory of Neurobiology, NIH) for supplying the squid axons used in these studies, to Dr. Jonathan Scholey and Dr. Gregory Rogers for providing the bacterial cells expressing p50/dynamitin, to Dr. Thomas Südhof for the synapsin expression vector, and to Dr. David Begg, Dr. Daniel Branton, and Dr. Eugeni Vaisberg for antibodies. This work was supported by a grant to B. J. S. from the National Institutes of Health (NS-26846). V. M. was supported in part by a Neuromuscular Disease Research Fellowship from the Muscular Dystrophy Association. E. L. F. H. was supported by a grant from the National Institutes of Health (GM-48661) and an Established Investigator Award from the American Heart Association. W. S. was supported by a grant from the Austrian Science Foundation (FWF #P12868-GEN), and a grant from the Deutsche Forschungsgemeinschaft (WE 790/18-1). J. S. M. was supported by grants from the National Institutes of Health.
PY - 2001
Y1 - 2001
N2 - We reconstituted dynein-driven, dynactin-dependent vesicle transport using protein-free liposomes and soluble components from squid axoplasm. Dynein and dynactin, while necessary, are not the only essential cytosolic factors; axonal spectrin is also required. Spectrin is resident on axonal vesicles, and rebinds from cytosol to liposomes or proteolysed vesicles, concomitant with their dynein-dynactin-dependent motility. Binding of purified axonal spectrin to liposomes requires acidic phospholipids, as does motility. Using dominant negative spectrin polypeptides and a drug that releases PH domains from membranes, we show that spectrin is required for linking dynactin, and thereby dynein, to acidic phospholipids in the membrane. We verify this model in the context of liposomes, isolated axonal vesicles, and whole axoplasm. We conclude that spectrin has an essential role in retrograde axonal transport.
AB - We reconstituted dynein-driven, dynactin-dependent vesicle transport using protein-free liposomes and soluble components from squid axoplasm. Dynein and dynactin, while necessary, are not the only essential cytosolic factors; axonal spectrin is also required. Spectrin is resident on axonal vesicles, and rebinds from cytosol to liposomes or proteolysed vesicles, concomitant with their dynein-dynactin-dependent motility. Binding of purified axonal spectrin to liposomes requires acidic phospholipids, as does motility. Using dominant negative spectrin polypeptides and a drug that releases PH domains from membranes, we show that spectrin is required for linking dynactin, and thereby dynein, to acidic phospholipids in the membrane. We verify this model in the context of liposomes, isolated axonal vesicles, and whole axoplasm. We conclude that spectrin has an essential role in retrograde axonal transport.
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U2 - 10.1016/S1097-2765(01)00165-4
DO - 10.1016/S1097-2765(01)00165-4
M3 - Article
C2 - 11172722
AN - SCOPUS:0035104723
SN - 1097-2765
VL - 7
SP - 173
EP - 183
JO - Molecular cell
JF - Molecular cell
IS - 1
ER -