To ensure genome stability, the spindle assembly checkpoint (SAC) delays anaphase until all pairs of kinetochores are attached to the ends of microtubule fibers from opposite spindle poles. In addition, improper chromosome-spindle linkages are selectively destroyed before anaphase. Through genome editing and chemical genetics, we found that the SAC kinase Mps1 controls both responses. Through large-scale phosphoproteomics, we discovered novel Mps1-regulated substrates at the kinetochore, including the Ska complex (which helps the Ndc80 complex grip onto dynamic microtubule ends) and the coatomer-related RZZ complex (which integrates Mad1-Mad2 dependent SAC signaling and dynein-dependent transport).
In Aim 1, we investigate how Mps1 affects the synergistic interactions of the Ska and Ndc80 complexes on dynamic microtubules, both in cells and in reconstituted in vitro systems.
In Aim 2, we dissect how Mps1 activates the RZZ complex for Mad1-Mad2 recruitment, long-term SAC arrest, and structural expansion of unattached kinetochores.
In Aim 3, we develop a new chemical- genetic system for Aurora A, a mitotic kinase that remains difficult to study because current inhibitors with Aurora A selectivity in vitro nonetheless cross-inhibit Aurora B at bioactive concentrations in vivo. Together these studies will reveal how kinetochore structure, microtubule attachment, and SAC signaling evolve during mitosis, in order to maximize the probability of error-free chromosome segregation. Ultimately this information will empower development of therapeutic agents that target aneuploidy- associated diseases such as cancer.

Public Health Relevance

Errors in transmitting chromosomes to dividing cells are strongly linked to human diseases, including infertility, spontaneous miscarriages, birth defects, and malignancy. The studies in this project seek to understand how the process of chromosome transmission is controlled at the molecular level. This information is necessary to understand how errors in chromosome transmission arise in the course of disease, and to develop new therapies that exploit this trait as an ?Achilles heel? of most cancer cells, providing more effective and less toxic cures for this disorder.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM094972-15
Application #
9705880
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Melillo, Amanda A
Project Start
2005-07-01
Project End
2022-05-31
Budget Start
2019-06-01
Budget End
2020-05-31
Support Year
15
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Combes, Guillaume; Barysz, Helena; Garand, Chantal et al. (2018) Mps1 Phosphorylates Its N-Terminal Extension to Relieve Autoinhibition and Activate the Spindle Assembly Checkpoint. Curr Biol 28:872-883.e5
Maciejowski, John; Drechsler, Hauke; Grundner-Culemann, Kathrin et al. (2017) Mps1 Regulates Kinetochore-Microtubule Attachment Stability via the Ska Complex to Ensure Error-Free Chromosome Segregation. Dev Cell 41:143-156.e6
Jones, Mathew J K; Jallepalli, Prasad V (2016) Engineering and Functional Analysis of Mitotic Kinases Through Chemical Genetics. Methods Mol Biol 1413:349-63
Kim, Minhee; O'Rourke, Brian P; Soni, Rajesh Kumar et al. (2016) Promotion and Suppression of Centriole Duplication Are Catalytically Coupled through PLK4 to Ensure Centriole Homeostasis. Cell Rep 16:1195-1203
Pagan, Julia K; Marzio, Antonio; Jones, Mathew J K et al. (2015) Degradation of Cep68 and PCNT cleavage mediate Cep215 removal from the PCM to allow centriole separation, disengagement and licensing. Nat Cell Biol 17:31-43
Rahman, Sadia; Jones, Mathew J K; Jallepalli, Prasad V (2015) Cohesin recruits the Esco1 acetyltransferase genome wide to repress transcription and promote cohesion in somatic cells. Proc Natl Acad Sci U S A 112:11270-5
Chen, Yu-Hung; Jones, Mathew J K; Yin, Yandong et al. (2015) ATR-mediated phosphorylation of FANCI regulates dormant origin firing in response to replication stress. Mol Cell 58:323-38
Rodriguez-Bravo, Veronica; Maciejowski, John; Corona, Jennifer et al. (2014) Nuclear pores protect genome integrity by assembling a premitotic and Mad1-dependent anaphase inhibitor. Cell 156:1017-31
Jones, Mathew J K; Jallepalli, Prasad V (2012) Chromothripsis: chromosomes in crisis. Dev Cell 23:908-17
Oppermann, Felix S; Grundner-Culemann, Kathrin; Kumar, Chanchal et al. (2012) Combination of chemical genetics and phosphoproteomics for kinase signaling analysis enables confident identification of cellular downstream targets. Mol Cell Proteomics 11:O111.012351

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