We will investigate the controls for centriole duplication and how prolonged prometaphase blocks daughter cell proliferation. Since mitotic fidelity depends upon the cell containing just two centrosomes, centrosome duplication must be under tight numerical and temporal control. Extra centrosomes (centrosome amplification) at mitosis can lead to unequal chromosome distribution and consequent genomic instability which is a driving force in multi- step carcinogenesis.
In Aim 1 we test a model that can explain centrosome amplification after DNA damage, a well established but poorly understood phenomenon. Radiation and radiomimetic drugs are currently used to treat human tumors. Follow up radiation therapy can cause DNA damage in proliferating normal cells in the tumor region - particularly after surgery.
In Aim 2 we will investigate if and how geminin plays a role in enforcing the block to centrosome reduplication during S and G2 phases of the cell cycle. The centrosome intrinsic block to reduplication is of critical importance in preventing centrosome amplification.
In Aim 3 we test if targeting of Cdk2-cyclin E to the centrosome is required for centrosome duplication or if soluble pools of this kinase complex are sufficient to drive centrosome duplication. This explores the importance of local control of centrosome duplication and provides insight into how zygotes control centrosome duplication despite constitutively high cytoplasmic Cdk2-cyclin E activity.
In Aim 4 we will further characterize the basis for our discovery that prolonged prometaphase causes an irreversible block to daughter cell proliferation despite the normal division of the mother cell. This proliferation block can serve to handle mitotic defects due to environmental toxins that are never resolved but allow satisfaction of the mitotic checkpoint and consequent improper completion of mitosis. We will also explore the possibility that chemotherapeutic regimes using microtubule targeting drugs could lead to stem cells withdrawing from the cell cycle thereby compromising tissue regeneration, wound healing, and tissue maintenance.

Public Health Relevance

The research outlined in this proposal investigates the mechanisms used by the cell to prevent errors in cell division which compromise genomic integrity. Genomic instability, a hallmark of many human cancers, is a recognized driving force in the evolution of aggressive growth characteristics and resistance to chemotherapeutic agents in clinical use.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Deatherage, James F
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University of Massachusetts Medical School Worcester
Anatomy/Cell Biology
Schools of Medicine
United States
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Uetake, Yumi; Sluder, Greenfield (2018) Activation of the apoptotic pathway during prolonged prometaphase blocks daughter cell proliferation. Mol Biol Cell 29:2632-2643
Duronio, Robert J; O'Farrell, Patrick H; Sluder, Greenfield et al. (2017) Sophisticated lessons from simple organisms: appreciating the value of curiosity-driven research. Dis Model Mech 10:1381-1389
Sluder, Greenfield (2016) Using sea urchin gametes and zygotes to investigate centrosome duplication. Cilia 5:20
Lambrus, Bramwell G; Daggubati, Vikas; Uetake, Yumi et al. (2016) A USP28-53BP1-p53-p21 signaling axis arrests growth after centrosome loss or prolonged mitosis. J Cell Biol 214:143-53
Wu, Qiong; Madany, Pasil; Akech, Jacqueline et al. (2015) The SWI/SNF ATPases Are Required for Triple Negative Breast Cancer Cell Proliferation. J Cell Physiol 230:2683-94
Lambrus, Bramwell G; Uetake, Yumi; Clutario, Kevin M et al. (2015) p53 protects against genome instability following centriole duplication failure. J Cell Biol 210:63-77
Sluder, Greenfield (2014) One to only two: a short history of the centrosome and its duplication. Philos Trans R Soc Lond B Biol Sci 369:
Ward, C L; Boggio, K J; Johnson, B N et al. (2014) A loss of FUS/TLS function leads to impaired cellular proliferation. Cell Death Dis 5:e1572
Douthwright, Stephen; Sluder, Greenfield (2014) Link between DNA damage and centriole disengagement/reduplication in untransformed human cells. J Cell Physiol 229:1427-36
Sluder, Greenfield; Nordberg, Joshua J (2013) Microscope basics. Methods Cell Biol 114:1-10

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