Haploinsufficiency of human LIS1 (or PAFAHIB1) is responsible for the human neuronal migration defect lissencephaly. Recently, we found that LIS1 interacts with NUDEL, a homologue of the A. nidulans nuclear distribution mutant NudE. LIS1 and NUDEL together bind the cytoplasmic dynein heavy chain (CDHC) to regulate dynein motor function in non-neural cells. NUDEL is phosphorylated by Cdk5/p35, a complex essential for neuronal migration in mice, suggesting that the LIS1/NUDEL/CDHC complex is regulated by phosphorylation. Phosphorylated NUDEL binds to 14-3-3epsilon, a member of a large family of binding proteins that mediate subcellular localization or stability of phosphoproteins. These recent studies have provided a pathway through which LIS1 acts to regulate dynein motor function in non-neural cells. However, it is unknown if this pathway is critical for neuronal migration. It is tempting to speculate that the dysregulation of dynein motor function by the LIS1/NUDEL/CDHC complex is also responsible for the neuronal migration defects in mammals with reduced doses of LIS1, mice deficient for the Cdk5/p35 complex, and mice deficient for 14-3-3epsilon. We hypothesize that LIS1 forms a complex (LIS1/NUDEL/CDHC) that regulates dynein motor function during neuronal migration. The function of this complex is regulated by phosphorylation of NUDEL by Cdk5/p35 via binding and regulation of the cellular distribution and/or stability of NUDEL by 14-3-3epsilon. To address these hypotheses, we will utilize genetic and gene transfer approaches to modulate the levels of specific components of this pathway in mice or cells. The effects of these modifications on neuronal migration will be determined in vivo and in vitro using quantitative migration assays. To prove whether effects of these modifications on neuronal migration result from alterations in the LIS1/NUDEL/CDHC complex, we will employ cell biological and biochemical assays of cytoplasmic dynein function, including the cellular localization and biochemical interaction of members of this complex.
The specific aims of this application are: 1) determine whether LIS1 participates in neuronal migration by regulation of dynein motor function, 2) determine whether NUDEL participates with LIS1 to regulate dynein motor functions via Cdk5/p35 phosphorylation during neuronal migration; and 3) to determine if 14-3-3epsilon regulates the cellular distribution and/or stability of Cdk5-phosphorylated NUDEL during neuronal migration to regulate the activity of LIS1 and CDHC.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS041030-03
Application #
6771158
Study Section
Molecular, Cellular and Developmental Neurosciences 2 (MDCN)
Program Officer
Leblanc, Gabrielle G
Project Start
2002-03-01
Project End
2006-01-31
Budget Start
2004-02-01
Budget End
2005-01-31
Support Year
3
Fiscal Year
2004
Total Cost
$361,000
Indirect Cost
Name
University of California San Diego
Department
Pediatrics
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Moon, Hyang Mi; Youn, Yong Ha; Pemble, Hayley et al. (2014) LIS1 controls mitosis and mitotic spindle organization via the LIS1-NDEL1-dynein complex. Hum Mol Genet 23:449-66
Toba, Shiori; Tamura, Yasuhisa; Kumamoto, Kanako et al. (2013) Post-natal treatment by a blood-brain-barrier permeable calpain inhibitor, SNJ1945 rescued defective function in lissencephaly. Sci Rep 3:1224
Moon, Hyang Mi; Wynshaw-Boris, Anthony (2013) Cytoskeleton in action: lissencephaly, a neuronal migration disorder. Wiley Interdiscip Rev Dev Biol 2:229-45
Takitoh, Takako; Kumamoto, Kanako; Wang, Chen-Chi et al. (2012) Activation of Aurora-A is essential for neuronal migration via modulation of microtubule organization. J Neurosci 32:11050-66
Toba, Shiori; Hirotsune, Shinji (2012) A unique role of dynein and nud family proteins in corticogenesis. Neuropathology 32:432-9
Wynshaw-Boris, Anthony; Pramparo, Tiziano; Youn, Yong Ha et al. (2010) Lissencephaly: mechanistic insights from animal models and potential therapeutic strategies. Semin Cell Dev Biol 21:823-30
Yamada, Masami; Hirotsune, Shinji; Wynshaw-Boris, Anthony (2010) A novel strategy for therapeutic intervention for the genetic disease: preventing proteolytic cleavage using small chemical compound. Int J Biochem Cell Biol 42:1401-7
Pramparo, Tiziano; Youn, Yong Ha; Yingling, Jessica et al. (2010) Novel embryonic neuronal migration and proliferation defects in Dcx mutant mice are exacerbated by Lis1 reduction. J Neurosci 30:3002-12
Yamada, Masami; Toba, Shiori; Takitoh, Takako et al. (2010) mNUDC is required for plus-end-directed transport of cytoplasmic dynein and dynactins by kinesin-1. EMBO J 29:517-31
Yamada, Masami; Hirotsune, Shinji; Wynshaw-Boris, Anthony (2010) The essential role of LIS1, NDEL1 and Aurora-A in polarity formation and microtubule organization during neurogensis. Cell Adh Migr 4:180-4

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