Vallee, Ph.D. Professor of Pathology and Cell Biology
Our lab is interested in a variety of biological phenomena involving motor proteins, with a major emphasis on cytoplasmic dynein and kinesins, their roles in diverse basic cell functions, and their contributions to disease. We identified dynein as the motor for retrograde axonal transport. It is now known to play important roles in mitosis, cell migration, growth cone motility, virus transport, and many other aspects of neuronal and nonneuronal cell behavior. Cytoplasmic dynein and the kinesins are also involved in diverse diseases of the nervous system, including lissencephaly and microcephaly, in which motor protein-mediated defects in brain stem cell behavior play a major role. The same motor proteins are also involved in CMT, SMA and other motor neuron diseases. Current projects are focused on the role of motor proteins in motor neuron and brain developmental disease, brain stem cell behavior and regulation, with recent emphasis the role of primary cilia, and basic mechanisms involved in transport and mitosis, with recent emphasis on autophagy and SMA. Projects involve extensive high-resolution live imaging in cultured neuronal and nonneuronal cells, and in live brain tissue, as well as basic cell biology and protein chemistry.
One current project involves the role of cytoplasmic dynein and kinesins in the human brain developmental diseases lissencephaly (smooth brain) and microcephaly. Lissencephaly arises from mutations in the dynein regulatory gene LIS1. A major approach to understanding how altered LIS1 expression alters brain development involves utero electroporation of neural progenitor cells in embryonic rat brain with shRNAs and cDNAs encoding mutant polypeptides and subcellular markers for microtubules, nuclei, centrosomes, and other structures. shRNAs are used to interfere with expression of LIS1, cytoplasmic dynein, and a variety of additional regulators, including NudE, NudEL, BicD2, and CENP-F, as well as the kinesin Kif1a. Fluorescent fusion protein markers are coexpressed to monitor cellular and subcellular behavior to understand the mechanisms underlying normal and abnormal neuronal migration, morphogenesis, and proliferation. These studies have led to models for how LIS1 and NudE mutations cause lissencephaly and microcephaly, and how forces generated by cytoplasmic dynein in developing neurons contribute to neural progenitor cell migration and division.
A relatively recent of these studies has been on the earliest stages of neurogenesis in the CNS, addressing the unusual behavior of the radial glial progenitor (RGP) cells. These highly elongated cells span the developing neocortex from the ventricles to the outer brain surface, and serve as guides for migrating neurons. In addition, the RGP cells are highly proliferative and give rise to most neurons and glial cells in the developing brain, and to adult stem cells. We have found that cytoplasmic dynein and its regulatory genes control the apical direction of nuclear migration, whereas Kif1a controls the basal direction. We have identified a novel checkpoint mechanism which prevents apically migrating nuclei from entering mitosis until they reach the surface of the ventricle. We also recently found Kif1a to control mitotic spindle orientation in the RGP cells, and thereby the relative production of neurons throughout development.
The lab has also investigated the molecular mechanisms by which LIS1 and other regulators control dynein motor function, as well as its subcellular targeting. We have found that LIS1, aided by NudE, binds to the dynein motor domain specifically during its powerstroke, stabilizing the interaction of dynein with microtubules during this phase of the crossbridge cycle (with lab of S. Gross). The result is a substantial increase in total forces generated by groups of dynein molecules, e. g. those associated with nuclei in the developing brain. We are also dissecting the molecular mechanisms involved in regulation of dynein by dynactin, NudE, NudEL, and several other factors, both in vitro and in live mitotic and neuronal cells. A recent focus of our work is the role of NudE in linking dynein and LIS1 to subcellular forms of cargo, and the cell cycle mechanisms controlling NudE and Kif1a behavior during mitosis in general and in brain development. Other work in the lab seeks to understand the mechanisms used by viruses to highjack dynein and other motor proteins for use in transport to the nucleus. We have used biochemical methods to identify motor proteins and their specific subunits recruited to adenovirus as a model organism, and the capsid components responsible for these interactions. We recently found that PKA activation by adenovirus infection results in specific phosphorylation of the dynein LIC1 subunit. This modification is essential for dynein recruitment by the virus, but also displaces dynein from lysosomes and late endosomes. The result is dispersal of these organelles to the cell periphery, an apparent host defense mechanism which the virus has evolved to bypass. We are also using high temporal resolution and high spatial resolution particle tracking analysis to monitor the behavior of fluorescently tagged virus in infected cells, to determine the role of kinesins as well as cytoplasmic dynein in the overall infection process.
Finally, we are investigating a number of genes involved in axonal transport and polarization of cultured neurons. This work involves high temporal resolution tacking of vesicular organelles, including autophagosomes, lysosomes, mitochondria, and other subcellular components important for basic neuronal function, homeostasis, and disease. One particular interest is the effect of human motor neuron disease-causing mutations on axonal transport.
Fig. 1: Neuronal migration in live rat brain slices. Centrosome moves continuously, followed by very discontinuous nuclear movements. Cytoplasmic dynein RNAi (right) partially inhibits centrosome movement, but arrests the nucleus (which moves out of the focal plane). Centrosomes labeled with RFP centrin (shown in green); nucleus labeled with histone H1 (shown in red). From Tsai, J.-W., et al., 2007.
Fig. 2: Behavior of neuronal precursor cell microtubules in live brain slices. Plus ends of growing microtubules are labeled with GFP-EB3 (green); centrosomes with RFP centrin (red). Movie at left focuses on microtubules in migratory process; movie at right includes microtubules in cell body region. From Tsai, J.-W., et al., 2007.
Fig. 3: Metaphase kinetochores in control (left) and cytoplasmic dynein defective (right) HeLa cells. Dynein inhibition disrupts normal oscillations of paired kinetochores, consistent with abnormal microtubule attachment. Kinetochores labeled with GFP-CENP-A; cells were arrested in metaphase with Mg132. (From Varma, D., et al., 2008).
Tsai, J.W., Bremner, K.H. , and Vallee, R.B. (2007). Dual subcellular roles for LIS1 and dynein in radial neuronal migration in live brain tissue. Nat Neurosci. 10:970-9.
Wang, X., Tsai, J. W., Imai, J. H., Lian, W. N., Vallee, R. B., and Shi, S. H. (2009). Asymmetric centrosome inheritance maintains neural progenitors in the neocortex. Nature 461, 947-955.
Vallee, R. B., Seale, G. E., and Tsai, J.-W. (2009) Emerging Roles for Myosin II and Cytoplasmic Dynein in Migrating Neurons and Growth Cones. Trends Cell Biol.19:347-355.
Bremner, K. H., Scherer, J., Yi, J., Vershinin, M., Gross, S. P., and Vallee, R. B. (2009). Adenovirus transport via direct interaction of cytoplasmic dynein with the viral capsid hexon subunit. Cell Host Microbe 6, 523-535.
McKenney, R. J.,* Vershinin, M.,* Vallee R. B.,+ and Gross, S. P.+ (2010) LIS1 and NudE induce a persistent dynein force-producing state. Cell 141:304-314. (*,+ Equal authorship.)
*Ori-McKenney, K. M.., *Xu, J., +Gross, S. P., and +Vallee, R. B. (2010) A Cytoplasmic Dynein Tail Mutation Impairs Motor Processivity Nature Cell Biol. 12:1228-1234.
*Tsai, J.-T., *Lian, W.-N., Kemal, S., Kriegstein, A., and Vallee, R. B. (2010) Kinesin 3 and Cytoplasmic Ddynein Mediate Interkinetic Nuclear Migration in Neural Stem Cells. Nature Neurosci. 13:1463-1471.
Ori-McKenney, K. M. and Vallee, R. B. (2011) Neuronal Migration Defects in the Loa Dynein Mutant Mouse. Neural Dev. 6:26.
Yi,J, Ori-McKenney, K. M., McKenney, R. J., Vershinin, M., Gross, S. P., and Vallee, R. B. (2011). High resolution imaging reveals indirect coordination of opposite motors and LIS1 role in high-load axonal transport. J. Cell Biol. 195:193-201.
McKenney R. J., Weil, S. J., Scherer, J. and Vallee, R. B. (2011) Mutually Exclusive Cytoplasmic Ddynein Regulation by NudE-LIS1 and Dynactin. J. Biol. Chem. 286:39615-39622.
Vallee, R. B., McKenney, R. J., and Ori-McKenney, K. M. (2012) Multiple Modes of Cytoplasmic Dynein Regulation. Nature Cell Biol, 14:224-230.
Fiorillo, C., Moro, F., Yi, J., Weil, S., Brisca, G., Astrea, G. Severino, M, Romano, A., Battini, R., Rossi A., Minetti, C., Bruno, C., Santorelli, F., and Vallee, R. B. (2013) Novel Dynein DYNC1H1 Neck and Motor Domain Mutations Link Distal SMA and Abnormal Cortical Development. Human Mut, 35:298-302.
Hu, D. J., Baffet, A. D., Nayak, T., Akhmanova, A., Doye, V., and Vallee, R. B. (2013) Dynein Recruitment to Nuclear Pores Activates Apical Nuclear Migration and Mitotic Entry in Brain Progenitor Cells. Cell 154, 1300–1313.
Scherer, J., Yi, J., Vallee, R. B. (2014) PKA-dependent Dynein Cargo Switching from Lysosomes to Adenovirus: a Novel Form of Host-Virus Competition. J.Cell Biol, 205:163-177.
Tripathy, S.*, Weil, S. J.*, Chen, C., Anand, P., Vallee, R. B.+, and Gross, S.+ (2014) Autoregulatory Mechanism for Dynactin Control of Processive and Diffusive Dynein Transport. Nature Cell Biol, 16:1192-1201.
Scherer, J., and Vallee, R. B. (2015) Conformational Changes in Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment. J. Virol. 89:1013-23.
Nicholas, M. P.,* Hook, P.,* Brenner, S., Lazar, C., Vallee, R. B., + and Gennerich, A.+ (2015) Control of cytoplasmic dynein force production and processivity by its C-terminal domain. Nature Commun. 6:6206.
Baffet, A. D., Hu, D. J., and Vallee, R. B. (2013) Cdk1 Activates Pre-Mitotic Nuclear Envelope Dynein Recruitment and Apical Nuclear Migration in Neural Stem cells. Dev. Cell 6:703-716.
Taylor, S. P,* Dantas, T. J.,* Duran, I., Wu, S., Lachman, R., Nelson, S., Cohn, D., Vallee, R. B., and Krakow, D. (2015) Mutations in DYNC2LI1 disrupt cilia function and cause short rib polydactyly syndrome. Nature Commun. 6:7092
Carabalona, A., Hu, D. J., and Vallee, R. B (2016) KIF1A Inhibition Immortalizes Brain Stem Cells but Blocks BDNF-mediated Neuronal Migration. Nature Neurosci.19:261-271.
(*Co-first and +Co-senior authors)
Research Assistant / Research Associate, Columbia University
Basic biomedical research in cell, molecular, and neurobiology. Entry or advanced level independent research on motor proteins involved in axonal transport, mitosis, brain developmental disease, neurodegeneration, virus transport, and related areas. B.A., M. A., Ph. D., or equivalent, and research experience required, especially in protein expression and purification, light microscopy, or molecular biology. Submit resume to firstname.lastname@example.org. Columbia University is an equal opportunity employer.
Honors and Awards
1996 NIH Merit Award
Council Delegate, AAAS
1999-2001 H. Arthur Smith
Chair in Cancer Research
Committees , Council, and Professional Society
1989 Associate Editor, Cell Motil.
1986, 1991, 1998 Series Editor, Methods in
Enzymology, Vol. 134, 196, and 298
Advisory Committee on Cell and Molecular Biology
1989-1994 Editorial Board, Journal of Biological