? Phosphoserine/threonine-binding domains play critical roles in controlling multiple aspects of cell proliferation, including cell cycle progression and the cellular response to DNA damage. The current list of pSer/pThr-binding domains includes 14-3-3 proteins, FHA domains, WW domains, WD40 repeats of F-box proteins, tandem BRCT domains and the Polo-box domains of Polo-like kinases. The long-term goal of our laboratory is to identify and characterize these domains with a focus on identifying their physiological ligands, determining the structural basis for their pSer/Thr-motif recognition, and elucidating the molecular basis for their functions in cell cycle control within complex protein kinase signaling networks. Polo-like kinases are essential during multiple stages of the eukaryotic cell cycle, including many of the events that occur during M-phase, as well as in the DNA damage response. Despite their importance, details about how Polo-like kinase activity is regulated, and the identity of their substrates are poorly understood. In this proposal we use a combination of biochemical, structural and cell biological techniques to determine the function of the invariant pSer/pThr-binding Polo-box domain in regulating the activation of, and the substrate selection and phosphorylation by Polo-like kinases. In the process, we will identify many new Polo-like kinase ligands and substrates that may be responsible for the pleiotropic role these kinases play in the cell. Since Polo-like kinases are upregulated in many types of human cancer, and since their experimental down-regulation results in decreased cell proliferation and tumor regression, the results of our experiments should determine whether the Polo-box domain is a good target for novel anti-cancer drug design. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM060594-09
Application #
7283024
Study Section
Biochemistry Study Section (BIO)
Program Officer
Smith, Ward
Project Start
2000-09-01
Project End
2009-08-31
Budget Start
2007-09-01
Budget End
2009-08-31
Support Year
9
Fiscal Year
2007
Total Cost
$282,154
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Internal Medicine/Medicine
Type
Schools of Arts and Sciences
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02139
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Morandell, Sandra; Reinhardt, H Christian; Cannell, Ian G et al. (2013) A reversible gene-targeting strategy identifies synthetic lethal interactions between MK2 and p53 in the DNA damage response in vivo. Cell Rep 5:868-77
Tentner, Andrea R; Lee, Michael J; Ostheimer, Gerry J et al. (2012) Combined experimental and computational analysis of DNA damage signaling reveals context-dependent roles for Erk in apoptosis and G1/S arrest after genotoxic stress. Mol Syst Biol 8:568
Liu, Fa; Park, Jung-Eun; Qian, Wen-Jian et al. (2012) Peptoid-Peptide hybrid ligands targeting the polo box domain of polo-like kinase 1. Chembiochem 13:1291-6
Liu, Fa; Park, Jung-Eun; Qian, Wen-Jian et al. (2012) Identification of high affinity polo-like kinase 1 (Plk1) polo-box domain binding peptides using oxime-based diversification. ACS Chem Biol 7:805-10
Liu, Fa; Park, Jung-Eun; Qian, Wen-Jian et al. (2011) Serendipitous alkylation of a Plk1 ligand uncovers a new binding channel. Nat Chem Biol 7:595-601
Gardino, Alexandra K; Yaffe, Michael B (2011) 14-3-3 proteins as signaling integration points for cell cycle control and apoptosis. Semin Cell Dev Biol 22:688-95
Hegemann, Björn; Hutchins, James R A; Hudecz, Otto et al. (2011) Systematic phosphorylation analysis of human mitotic protein complexes. Sci Signal 4:rs12
Alexander, Jes; Lim, Daniel; Joughin, Brian A et al. (2011) Spatial exclusivity combined with positive and negative selection of phosphorylation motifs is the basis for context-dependent mitotic signaling. Sci Signal 4:ra42
Wen, Jian; Wilker, Erik W; Yaffe, Michael B et al. (2010) Microfluidic preparative free-flow isoelectric focusing: system optimization for protein complex separation. Anal Chem 82:1253-60

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