Centrosomes are best known for their ability to organize microtubules to form interphase arrays and mitotic spindles. Recent evidence suggests that centrosomes are involved hi cytokinesis and cell cycle progression. However, little is known about the centrosome proteins that contribute to these processes. In the previous funding period, we examined the molecular basis for these under-explored centrosome functions. We identified a novel centrosome protein called centriolin, which is essential for cytokinesis in mammalian cells. Centriolin interacts with proteins implicated or involved in cytokinesis including snapin (SNARE), sec 15 (exocyst), MKLP-1 (centralspindlin) and a novel GTPase activating protein for Rho GTPases. Many of these proteins localize to the midbody in dividing cells and induce cytokinesis defects when downregulated. Based on these and other data, we propose a model in which centriolin serves as a scaffold protein at the midbody to coordinate vesicle fusion, microtubule depolymerization, furrow ingression and cell separation late in cytokinesis. Defective cytokinesis induced by centriolin downregulation was followed by Gl arrest. We unexpectedly found that many centrosome proteins induced GI arrest when downregulated. However, the arrest did not correlate with defects in cytokinesis or other centrosome functions, or in centrosome structure or composition. We postulate that cell cycle arrest is triggered by a checkpoint that monitors defects in a common function not yet identified, or 'centrosome damage' induced by reduction of individual proteins at centrosomes. GI arrest requires p53 and p38 MAP kinase and induces recruitment of activated p53 and p38 to centrosomes. Based on these observations, we propose two specific aims.
In Aim 1 we will determine the role centriolin and associated proteins hi cytokinesis. More specifically, we will test whether centriolin anchors these proteins at the midbody and if a complex of these proteins controls vesicle fusion, microtubule organization and cell separation at the midbody.
Aim 2 focuses on the role of centrosome proteins in cell cycle progression and checkpoint activation. We will determine the mechanism of GI arrest and identify the signal transduction pathway that connects centrosome proteins to cell cycle arrest.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM051994-10A1
Application #
6918208
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Deatherage, James F
Project Start
1995-08-01
Project End
2009-03-31
Budget Start
2005-04-01
Budget End
2006-03-31
Support Year
10
Fiscal Year
2005
Total Cost
$382,709
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Hung, Hui-Fang; Hehnly, Heidi; Doxsey, Stephen (2016) The Mother Centriole Appendage Protein Cenexin Modulates Lumen Formation through Spindle Orientation. Curr Biol 26:793-801
Vertii, Anastassiia; Bright, Alison; Delaval, Benedicte et al. (2015) New frontiers: discovering cilia-independent functions of cilia proteins. EMBO Rep 16:1275-87
Vertii, Anastassiia; Zimmerman, Wendy; Ivshina, Maria et al. (2015) Centrosome-intrinsic mechanisms modulate centrosome integrity during fever. Mol Biol Cell 26:3451-63
Hung, Hui-Fang; Hehnly, Heidi; Doxsey, Stephen (2015) Methods to analyze novel liaisons between endosomes and centrosomes. Methods Cell Biol 130:47-58
de Souza, Edmarcia Elisa; Hehnly, Heidi; Perez, Arina Marina et al. (2015) Human Nek7-interactor RGS2 is required for mitotic spindle organization. Cell Cycle 14:656-67
Hehnly, Heidi; Doxsey, Stephen (2014) Rab11 endosomes contribute to mitotic spindle organization and orientation. Dev Cell 28:497-507
Chen, Chun-Ting; Hehnly, Heidi; Yu, Qing et al. (2014) A unique set of centrosome proteins requires pericentrin for spindle-pole localization and spindle orientation. Curr Biol 24:2327-2334
Chen, Chun-Ting; Ettinger, Andreas W; Huttner, Wieland B et al. (2013) Resurrecting remnants: the lives of post-mitotic midbodies. Trends Cell Biol 23:118-28
Kuo, Tse-Chun; Doxsey, Stephen (2012) Fates and roles of post-mitotic midbodies beyond cytokinesis. Cell Cycle 11:7-8
Foraker, Amy B; Camus, St├ęphane M; Evans, Timothy M et al. (2012) Clathrin promotes centrosome integrity in early mitosis through stabilization of centrosomal ch-TOG. J Cell Biol 198:591-605

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