Mammalian SCF (Skp1/Cul1/F-box protein) ubiquitin ligases are critical for the activation and attenuation of many cellular processes. They control complex molecular machines by directing the proteolysis of important regulatory elements in a precise, rapid, and localized manner. During the initial years of GM57587, we focused on the regulation of cyclin-dependent kinases (CDKs), which play pivotal roles in cell cycle control, and defined the key steps leading to the degradation of the CDK inhibitors p21 and p27 via the F-box protein Skp2. Moreover, we found that APC/CCdh1, an SCF-like ligase, controls Skp2 stability. We also demonstrated that ?TrCP, another F-box protein, allows precise regulation of the critical mitosis regulator Cdk1 by targeting Cdc25A, Claspin, Rest, and Emi1 for degradation. Finally, we found that three substrates that are targeted by SCF ubiquitn ligases in S and G2 are degraded via APC/C during different phases of the cell cycle (p21 in M, and Cdc25A and Claspin in G1). To broaden our understanding beyond CDK-centric roles of SCF complexes, we used unbiased screens and found that SCF, in addition to controlling the cell cycle, monitors and regulates multiple, seemingly disparate, cellular pathways, linking cell cycle control to protein synthesis, ribosomal biogenesis, cell survival, DNA-damage checkpoints, and the circadian clock. For example, we found that: the translation inhibitor Pdcd4 and the pro-apoptotic protein BimEL are degraded in a ?TrCP-dependent manner in response to growth and survival factors;Fbxl10 and Fbxl11 contribute to epigenetic regulation;Cyclin F/Fbxo1 prevents centrosome overduplication by targeting CP110 for degradation;and Fbxl3 is required to reset the circadian clock by promoting the proteolysis of the transcriptional repressors Cry1 and Cry2. We now propose a project exploring the integration of SCF-controlled cell cycle networks with DNA replication and DNA damage response. Via proteomic screens, we have identified novel putative SCF and APC/C substrates involved in these processes. We will characterize the mechanism and regulation of the degradation of these potential substrates in the context of DNA replication control (Aim 1), recovery from genotoxic stress (Aims 1 and 2), and mitosis (Aim 3).

Public Health Relevance

Cells depend on the proper functioning of an ensemble of networked, molecular machines to control diverse processes, ranging from cell proliferation to cell death to differentiation. The ubiquitin system can rapidly degrade the modular regulatory components of these machines, contributing to the precise operation and synchronization of complex cellular processes. Given its critical role, the ubiquitin system is often deregulated in cancer cells. Thus, it is anticipated that the results of the proposed studies will have an impact on both basic science and cancer biology.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057587-14
Application #
8242014
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Hamlet, Michelle R
Project Start
1998-09-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
14
Fiscal Year
2012
Total Cost
$456,031
Indirect Cost
$186,190
Name
New York University
Department
Pathology
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
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Skaar, Jeffrey R; Pagan, Julia K; Pagano, Michele (2014) SCF ubiquitin ligase-targeted therapies. Nat Rev Drug Discov 13:889-903
Bassermann, Florian; Eichner, Ruth; Pagano, Michele (2014) The ubiquitin proteasome system - implications for cell cycle control and the targeted treatment of cancer. Biochim Biophys Acta 1843:150-62
Kanarek, Naama; Grivennikov, Sergei I; Leshets, Michael et al. (2014) Critical role for IL-1β in DNA damage-induced mucositis. Proc Natl Acad Sci U S A 111:E702-11

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