Successful cell division requires exquisite spatial and temporal control of the cytoskeleton such that the ingressing cleavage furrow forms between the separating sisters chromatids. Contractile ring assembly occurs in the cell cortex, the cytoplasmic region juxtaposed to the plasma membrane. The mechanism by which the master regulator of this process, the small GTPase RhoA, is activated at the equatorial cell cortex is not well understood. New evidence has been obtained that reveals a previously unidentified inhibitory mechanism. Preliminary results suggest that characterization of this inhibitory mechanism will provide insights into how exchange factors promote RhoA activation. These results and evidence from a variety of systems suggest a hypothesis in which plasma membrane targeting of guanine nucleotide exchange factors is a key step in activating GTPase pathways and that, once active, the primary function that GTPases serve is to recruit effectors to the appropriate membrane environment. This hypothesis will be directly tested in the context of cytokinesis using recently developed optogenetic tools that affords control of protein localization and function in living cells using light. Lastly, experiments will be performed to uncover the molecular function of a novel GYF-domain containing protein that regulates RhoA-dependent cortical contractility.

Public Health Relevance

Cytokinesis, the generation of two daughter cells from a single progenitor, is fundamental to growth, development and homeostasis and is a preeminent example of a spatiotemporally controlled cellular program regulated by a GTPase, namely RhoA. The principles of RhoA regulation, and the downstream mechanisms through which RhoA controls cytokinesis, may be generalizable to other GTPases, including those that regulate cell growth, migration and membrane trafficking. This proposal utilizes an innovative optogenetic technique recently developed in the applicant's laboratory which will be used to investigate the mechanism of cytokinesis.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Cellular Signaling and Regulatory Systems Study Section (CSRS)
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Gindhart, Joseph G
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University of Chicago
Schools of Medicine
United States
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Wagner, Elizabeth; Glotzer, Michael (2016) Local RhoA activation induces cytokinetic furrows independent of spindle position and cell cycle stage. J Cell Biol 213:641-9
Basant, Angika; Lekomtsev, Sergey; Tse, Yu Chung et al. (2015) Aurora B kinase promotes cytokinesis by inducing centralspindlin oligomers that associate with the plasma membrane. Dev Cell 33:204-15
Zhang, Donglei; Glotzer, Michael (2015) Cytokinesis: Placing the Furrow in Context. Curr Biol 25:R1183-5
Zhang, Donglei; Glotzer, Michael (2015) The RhoGAP activity of CYK-4/MgcRacGAP functions non-canonically by promoting RhoA activation during cytokinesis. Elife 4:
White, Erin A; Raghuraman, Hariharasundaram; Perozo, Eduardo et al. (2013) Binding of the CYK-4 subunit of the centralspindlin complex induces a large scale conformational change in the kinesin subunit. J Biol Chem 288:19785-95
White, Erin A; Glotzer, Michael (2012) Centralspindlin: at the heart of cytokinesis. Cytoskeleton (Hoboken) 69:882-92
Tse, Yu Chung; Werner, Michael; Longhini, Katrina M et al. (2012) RhoA activation during polarization and cytokinesis of the early Caenorhabditis elegans embryo is differentially dependent on NOP-1 and CYK-4. Mol Biol Cell 23:4020-31
Loria, Andy; Longhini, Katrina M; Glotzer, Michael (2012) The RhoGAP domain of CYK-4 has an essential role in RhoA activation. Curr Biol 22:213-9
Tse, Yu Chung; Piekny, Alisa; Glotzer, Michael (2011) Anillin promotes astral microtubule-directed cortical myosin polarization. Mol Biol Cell 22:3165-75
Afshar, Katayoun; Werner, Michael E; Tse, Yu Chung et al. (2010) Regulation of cortical contractility and spindle positioning by the protein phosphatase 6 PPH-6 in one-cell stage C. elegans embryos. Development 137:237-47

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