Microtubules are dynamic structures that are required for several vital processes in eukaryotic cells including mitosis, the establishment and maintenance of cell shape and cell motility. The overall objective of our work is to determine the mechanism(s) by which microtubules are remodelled as cells traverse the cell cycle and locomote and how microtubules contribute to efficient cellular motility. A key feature of our experimental approach is the analysis of microtubules in the living cell. To accomplish this we will use quantitative digital fluorescence microscopy, photoactivation of caged tubulin, fluorescence analog cytochemistry and photobleaching of GFP- tagged proteins. The major aims are to determine the dynamic properties of the minus-ends of centrosomal microtubules; measure nucleation and release of centrosomal microtubules in motile and non-motile cells; determine the mechanism responsible for microtubule transport; determine if centrosome reorientation occurs during motion of fibroblast-like cells and during cell migration into a wounded monolayer; and determine the regulatory pathways that govern microtubule turnover in motile cells. Microtubule dynamics and remodelling will be studied in highly motile keratocytes and in two model systems for mammalian cell motility: migration into a wound and fibroblast-like motion in response to growth factors. The results will elucidate the mechanisms of microtubule turnover in live cells and contribute to our understanding of the role of microtubules in cell polarity and motility. Understanding these processes in normal cells is an important first step towards analysis of cancer cells, where such processes are aberrant.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM059057-03
Application #
6525510
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Deatherage, James F
Project Start
2000-08-01
Project End
2004-07-31
Budget Start
2002-08-01
Budget End
2003-07-31
Support Year
3
Fiscal Year
2002
Total Cost
$191,875
Indirect Cost
Name
University of Massachusetts Amherst
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
153223151
City
Amherst
State
MA
Country
United States
Zip Code
01003
Balchand, Sai K; Mann, Barbara J; Wadsworth, Patricia (2016) Using Fluorescence Microscopy to Study Mitosis. Methods Mol Biol 1413:3-14
Wadsworth, Pat (2015) TPX2. Curr Biol 25:R1156-8
Collins, Elizabeth; Mann, Barbara J; Wadsworth, Patricia (2014) Eg5 restricts anaphase B spindle elongation in mammalian cells. Cytoskeleton (Hoboken) 71:136-44
Wadsworth, Patricia; Lee, Wei-Lih; Murata, Takashi et al. (2011) Variations on theme: spindle assembly in diverse cells. Protoplasma 248:439-46
Ma, Nan; Titus, Janel; Gable, Alyssa et al. (2011) TPX2 regulates the localization and activity of Eg5 in the mammalian mitotic spindle. J Cell Biol 195:87-98
Ferenz, Nick P; Gable, Alyssa; Wadsworth, Pat (2010) Mitotic functions of kinesin-5. Semin Cell Dev Biol 21:255-9
Ferenz, Nick P; Ma, Nan; Lee, Wei-Lih et al. (2010) Imaging protein dynamics in live mitotic cells. Methods 51:193-6
Ma, Nan; Tulu, U S; Ferenz, Nick P et al. (2010) Poleward transport of TPX2 in the mammalian mitotic spindle requires dynein, Eg5, and microtubule flux. Mol Biol Cell 21:979-88
Tulu, U Serdar; Ferenz, Nick P; Wadsworth, Patricia (2010) Photoactivatable green fluorescent protein-tubulin. Methods Cell Biol 97:81-90
Miquelard-Garnier, Guillaume; Zimberlin, Jessica A; Sikora, Christian B et al. (2010) Polymer microlenses for quantifying cell sheet mechanics. Soft Matter 6:398-403

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