In all eukaryotes, entry into S phase and mitosis is triggered by protein kinases, whose activity depends on regulatory cyclin subunits (cyclin-dependent kinases; Cdks). Different types of Cdks initiate different events of the cell cycle. It is therefore critically important that their activities are restricted to the appropriate cell cycle stages. Ubiquitin-dependent proteolysis of mitotic cyclins plays a key role in restricting the activity of the mitotic Cdks. Mitotic cyclin proteolysis inactivates mitotic Cdks at the end of mitosis and prevents their accumulation throughout G1. During S phase and early mitosis, mitotic cyclin proteolysis is inhibited, allowing mitotic Cdks to accumulate. The goals of this proposal are: (1) to identify the pathway(s) of activation of mitotic cyclin proteolysis; (2) to determine the role of Cdks in repression of mitotic cyclin proteolysis; and (3) to identify genes required for repression of mitotic cyclin proteolysis during S-phase and early mitosis. To characterize the pathway(s) of activation of proteolysis, genes will be identified specifically defective in this process. These analyses will utilize assays for activation of proteolysis and maintenance of proteolysis, recently developed by Dr. Amon. Using these assays, the putative tyrosine phosphatase Cdc14 has already been identified as being specifically required for activation but dispensable for maintenance of mitotic cyclin proteolysis. The role of CDC14 in activation of mitotic cyclin proteolysis will be characterized in detail. Two approaches will be taken to investigate the role of Cdks in repression of mitotic cyclin proteolysis during S phase and early mitosis. (1) Biochemical experiments will address whether Cdk phosphorylation directly inhibits the proteolysis process. (2) A genetic approach will identify genes required for repression of mitotic cyclin proteolysis during S phase and early mitosis. Uncontrolled cyclin expression can be lethal. It has been linked to cancerous transformation: cyclin overexpression is a prominent feature of certain tumors including breast cancer. Identifying and characterizing the regulatory networks, such as mitotic cyclin proteolysis, that restrict the activity of these proteins will open avenues towards the development of cancer diagnostics and therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29GM056800-02
Application #
2771109
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1997-09-30
Project End
1999-01-31
Budget Start
1998-09-01
Budget End
1999-01-31
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
076580745
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Sheltzer, Jason M; Ko, Julie H; Replogle, John M et al. (2017) Single-chromosome Gains Commonly Function as Tumor Suppressors. Cancer Cell 31:240-255
Torres, Eduardo M; Springer, Michael; Amon, Angelika (2016) No current evidence for widespread dosage compensation in S. cerevisiae. Elife 5:e10996
Dodgson, Stacie E; Kim, Sharon; Costanzo, Michael et al. (2016) Chromosome-Specific and Global Effects of Aneuploidy in Saccharomyces cerevisiae. Genetics 202:1395-409
Knouse, Kristin A; Wu, Jie; Amon, Angelika (2016) Assessment of megabase-scale somatic copy number variation using single-cell sequencing. Genome Res 26:376-84
Blank, Heidi M; Sheltzer, Jason M; Meehl, Colleen M et al. (2015) Mitotic entry in the presence of DNA damage is a widespread property of aneuploidy in yeast. Mol Biol Cell 26:1440-51
Pfau, Sarah J; Amon, Angelika (2015) A System to Study Aneuploidy In Vivo. Cold Spring Harb Symp Quant Biol 80:93-101
Santaguida, Stefano; Amon, Angelika (2015) Aneuploidy triggers a TFEB-mediated lysosomal stress response. Autophagy 11:2383-4
Santaguida, Stefano; Amon, Angelika (2015) Short- and long-term effects of chromosome mis-segregation and aneuploidy. Nat Rev Mol Cell Biol 16:473-85
Bonney, Megan E; Moriya, Hisao; Amon, Angelika (2015) Aneuploid proliferation defects in yeast are not driven by copy number changes of a few dosage-sensitive genes. Genes Dev 29:898-903
Santaguida, Stefano; Vasile, Eliza; White, Eileen et al. (2015) Aneuploidy-induced cellular stresses limit autophagic degradation. Genes Dev 29:2010-21

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