Cell migration in higher organisms is essential for multiple physiological and pathophysiological processes, including embryonic development and immune responses. Alteration of cell motility in cancer cells is one of the most dangerous features of malignant tumors, as it builds up their invasive and metastatic potential. Polarized organization of microtubule arrays is essential for polarized cell motility. However, the principles of this regulation are not yet understood. It is generally assumed that microtubules in vertebrate cells are formed by the centrosome. We have recently demonstrated that a large number of microtubules originate from the Golgi apparatus (Efimov et al, 2007). We have identified two molecular players critical for this novel phenomenon: microtubule regulatory proteins CLASPs are required for the formation of Golgi-derived microtubules, and golgin GCC185 serves as an anchor for CLASPs at the trans-Golgi network (TGN) at the Golgi periphery. In sharp contrast to symmetric microtubule arrays organized by the centrosome, microtubules nucleated at the peripheral Golgi compartment are preferentially oriented toward the leading edge in motile cells. Preliminary data suggest that the migratory potential of cells lacking Golgi-originated microtubules is compromised. Within this proposal, we will test the hypothesis that asymmetric microtubule nucleation at the Golgi is critical for motile cell polarization. We will test whether Golgi-originated microtubules exert their effect on cell polarity via regulation of the actin cytoskeleton or directional post-Golgi transport to the cell front, or both. We will also address molecular mechanisms that regulate microtubule formation at the TGN.
Our specific aims are: 1. Determine the role of Golgi-derived microtubules in the cytoskeletal polarity of motile cells. 2. Determine the role of Golgi-derived microtubules in polarized Golgi trafficking in motile cells. 3. Determine if dynamic CLASP anchoring underlies the microtubule-organizing potential of the Golgi. PUBLIC HEALTH REVELANCE: Knowledge of the molecular events that underlie cell motility is critical for understanding major health-related processes, including embryonic morphogenesis, wound healing, the immune response and cancer invasiveness. As major anticancer therapies include microtubule-specific drugs, knowledge their potential targets is especially valuable. In this proposal, we will carry out research that will determine the role of a novel Golgi-derived asymmetric microtubule array in cell motility.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM078373-05
Application #
8241079
Study Section
Cell Structure and Function (CSF)
Program Officer
Deatherage, James F
Project Start
2008-04-01
Project End
2014-03-31
Budget Start
2012-04-01
Budget End
2014-03-31
Support Year
5
Fiscal Year
2012
Total Cost
$278,092
Indirect Cost
$96,773
Name
Vanderbilt University Medical Center
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Tonucci, Facundo M; Ferretti, Anabela; Almada, Evangelina et al. (2018) Microtubules regulate brush border formation. J Cell Physiol 233:1468-1480
Sanders, Anna A W M; Chang, Kevin; Zhu, Xiaodong et al. (2017) Nonrandom ?-TuNA-dependent spatial pattern of microtubule nucleation at the Golgi. Mol Biol Cell 28:3181-3192
Arnette, Christopher; Frye, Keyada; Kaverina, Irina (2016) Microtubule and Actin Interplay Drive Intracellular c-Src Trafficking. PLoS One 11:e0148996
Zhu, Xiaodong; Efimova, Nadia; Arnette, Christopher et al. (2016) Podosome dynamics and location in vascular smooth muscle cells require CLASP-dependent microtubule bending. Cytoskeleton (Hoboken) 73:300-15
Grimaldi, Ashley D; Zanic, Marija; Kaverina, Irina (2015) Encoding the microtubule structure: Allosteric interactions between the microtubule +TIP complex master regulators and TOG-domain proteins. Cell Cycle 14:1375-8
Sanders, Anna A W M; Kaverina, Irina (2015) Nucleation and Dynamics of Golgi-derived Microtubules. Front Neurosci 9:431
Tonucci, Facundo M; Hidalgo, Florencia; Ferretti, Anabela et al. (2015) Centrosomal AKAP350 and CIP4 act in concert to define the polarized localization of the centrosome and Golgi in migratory cells. J Cell Sci 128:3277-89
Zhu, Xiaodong; Hu, Ruiying; Brissova, Marcela et al. (2015) Microtubules Negatively Regulate Insulin Secretion in Pancreatic ? Cells. Dev Cell 34:656-68
Maki, Takahisa; Grimaldi, Ashley D; Fuchigami, Sotaro et al. (2015) CLASP2 Has Two Distinct TOG Domains That Contribute Differently to Microtubule Dynamics. J Mol Biol 427:2379-95
Cleghorn, Whitney M; Branch, Kevin M; Kook, Seunghyi et al. (2015) Arrestins regulate cell spreading and motility via focal adhesion dynamics. Mol Biol Cell 26:622-35

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