Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Cytoplasmic dynein is a microtubule motor protein involved in a very wide array of cellular functions, including vesicular transport, chromosome segregation, and cell migration. A single major form of cytoplasmic dynein is responsible for almost all aspects of these activities, but how it is adapted to such a diversity of functions at a broad range of subcellular sites remains a major question. Among the known dynein interactors, two complexes have emerged with prominent roles in dynein cargo binding and motor regulation, NudE-LIS1 and dynactin. We have recently reported NudE-LIS1 to have a novel and unique ability to interact with the dynein motor domain during its powerstroke and adapt the motor protein for high load functions. We also found LIS1-NudE to compete with dynactin for dynein binding, suggesting that the two complexes may serve in alternative regulatory roles. This proposal is to test this hypothesis and to bring to bear on dynactin the approaches we have successfully used to determine the functions and mechanism of action of LIS1 and NudE. Dynactin is known to enhance dynein processivity in vitro. However, its interaction with dynein has been difficult to control, hampering progress toward a complete understanding of its mechanochemical functions. Because dynactin is also important in dynein cargo recruitment, its specific in vivo role in motor regulation has also been difficult to define. Preliminary results have identified conditions controlling the dynein-dynactin interaction, and have revealed potent, long-range allosteric effects for dynactin fragments on dynein force production and processivity.
The Aims of this proposal are (1) to determine how the dynein-dynactin interaction is regulated and to produce and define cocomplexes for further analysis;(2) to determine the complete scope of regulatory functions for the dynactin complex, its major regulatory subunit p150Glued, and its subfragments to understand the underlying mechanisms for dynein regulation;and (3) to determine the specific roles of dynactin in dynein motor regulation in vivo by high resolution particle tracking and force analysis. These studies are of broad relevance for understanding basic mechanisms of cell behavior. In addition, they should shed important new light into the mechanisms underlying brain developmental disease, motor neuron degeneration, cell division, and other physiological and pathophysiological functions.

Public Health Relevance

Cytoplasmic dynein is a major microtubule motor protein involved in important aspects of intracellular transport, cell division, and cell migration. Dynactin is a prominent cytoplasmic dynein regulator. Mutations in cytoplasmic dynein and dynactin cause human motor and sensory neurodegeneration and defects in brain development. An understanding of dynein and dynactin function has important implications for managing these conditions and understanding and treating neoplastic diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM102347-01
Application #
8343218
Study Section
Synapses, Cytoskeleton and Trafficking Study Section (SYN)
Program Officer
Gindhart, Joseph G
Project Start
2012-09-28
Project End
2016-05-31
Budget Start
2012-09-28
Budget End
2013-05-31
Support Year
1
Fiscal Year
2012
Total Cost
$441,480
Indirect Cost
$141,480
Name
Columbia University (N.Y.)
Department
Pathology
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Wynne, Caitlin L; Vallee, Richard B (2018) Cdk1 phosphorylation of the dynein adapter Nde1 controls cargo binding from G2 to anaphase. J Cell Biol 217:3019-3029
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Yi, Julie; Khobrekar, Noopur V; Dantas, Tiago J et al. (2016) Imaging of motor-dependent transport in neuronal and nonneuronal cells at high spatial and temporal resolution. Methods Cell Biol 131:453-65
Reddy, Babu J N; Mattson, Michelle; Wynne, Caitlin L et al. (2016) Load-induced enhancement of Dynein force production by LIS1-NudE in vivo and in vitro. Nat Commun 7:12259
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Scherer, Julian; Vallee, Richard B (2015) Conformational changes in the adenovirus hexon subunit responsible for regulating cytoplasmic dynein recruitment. J Virol 89:1013-23
Baffet, Alexandre D; Hu, Daniel J; Vallee, Richard B (2015) Cdk1 Activates Pre-mitotic Nuclear Envelope Dynein Recruitment and Apical Nuclear Migration in Neural Stem Cells. Dev Cell 33:703-16
Taylor, S Paige; Dantas, Tiago J; Duran, Ivan et al. (2015) Mutations in DYNC2LI1 disrupt cilia function and cause short rib polydactyly syndrome. Nat Commun 6:7092
Nicholas, Matthew P; Höök, Peter; Brenner, Sibylle et al. (2015) Control of cytoplasmic dynein force production and processivity by its C-terminal domain. Nat Commun 6:6206
Tripathy, Suvranta K; Weil, Sarah J; Chen, Chen et al. (2014) Autoregulatory mechanism for dynactin control of processive and diffusive dynein transport. Nat Cell Biol 16:1192-201

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