Our overall goal is to determine the manner by which the peptidyl-prolyl isomerase, Pin1, acts as a negative regulator of mitosis and evaluate whether this protein may be an attractive target for development of an anti-cancer therapeutic agent. Pin1 was discovered as a protein that interacts with NIMA, a protein Ser/Thr Kinase that is essential for progression from G2 to mitosis in Aspergillus nidulans. Pin1 has been conserved from yeast to man and may be critical for mitotic progression. We cloned Pin1 from Asperigillus and Xenopus, and made antibodies to the protein. We showed that Pin1 will regulate entry into mitosis in Xenopus extracts where it interacts with key components of the p34cdc2 regulatory pathway such as the active forms of the cdc25 protein phosphatase and the plxl protein kinase as well as a number of other mitotic phosphoproteins recognized by the MPM2 antibody. We propose to use the Xenopus Egg/oocyte system to address key issues regarding the prolyl isomerase and protein binding of Pin1 and elucidate critical details of its role(s) as a cell cycle regulator. We will evaluate the tole of Pin1 in the timing of mitotic entry and in the operation of checkpoints governing the G2/M transition as well as examine if Pin1 plays a role in exit from mitosis. In parallel, we will determine if Pin1 is essential for mouse development and focus on its role in development, activation and/or proliferation of T lymphocytes by using the cre/lox system to disrupt the Pin1 gene in mice either globally or specifically in T lymphocytes. We will determine if phenotypic consequences of Pin1 deletion in either vertebrate system requires the prolyl isomerase activity. Completion of these studies will define the role(s) of Pin1 in growth and development, determine its critical cell cycle functions and clarify whether the Pin1 prolyl isomerase is a good candidate for anti-cancer drug development.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA082845-05S1
Application #
6895404
Study Section
Metabolic Pathology Study Section (MEP)
Program Officer
Spalholz, Barbara A
Project Start
1999-08-06
Project End
2005-05-31
Budget Start
2003-06-01
Budget End
2005-05-31
Support Year
5
Fiscal Year
2004
Total Cost
$52,206
Indirect Cost
Name
Duke University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Barberi, Theresa J; Dunkle, Alexis; He, You-Wen et al. (2012) The prolyl isomerase Pin1 modulates development of CD8+ cDC in mice. PLoS One 7:e29808
Teng, Brian L; Hacker, Kathryn E; Chen, Shufen et al. (2011) Tumor suppressive activity of prolyl isomerase Pin1 in renal cell carcinoma. Mol Oncol 5:465-74
Girardini, Javier E; Napoli, Marco; Piazza, Silvano et al. (2011) A Pin1/mutant p53 axis promotes aggressiveness in breast cancer. Cancer Cell 20:79-91
Swenson-Fields, Katherine I; Sandquist, Joshua C; Rossol-Allison, Jessica et al. (2008) MLK3 limits activated Galphaq signaling to Rho by binding to p63RhoGEF. Mol Cell 32:43-56
Sandquist, Joshua C; Means, Anthony R (2008) The C-terminal tail region of nonmuscle myosin II directs isoform-specific distribution in migrating cells. Mol Biol Cell 19:5156-67
Stanya, Kristopher J; Liu, Yu; Means, Anthony R et al. (2008) Cdk2 and Pin1 negatively regulate the transcriptional corepressor SMRT. J Cell Biol 183:49-61
Reineke, Erin L; Lam, Minh; Liu, Qing et al. (2008) Degradation of the tumor suppressor PML by Pin1 contributes to the cancer phenotype of breast cancer MDA-MB-231 cells. Mol Cell Biol 28:997-1006
Zita, M Moretto; Marchionni, Ivan; Bottos, Elisa et al. (2007) Post-phosphorylation prolyl isomerisation of gephyrin represents a mechanism to modulate glycine receptors function. EMBO J 26:1761-71
Sandquist, Joshua C; Swenson, Katherine I; Demali, Kris A et al. (2006) Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration. J Biol Chem 281:35873-83
Yeh, Elizabeth S; Lew, Brian O; Means, Anthony R (2006) The loss of PIN1 deregulates cyclin E and sensitizes mouse embryo fibroblasts to genomic instability. J Biol Chem 281:241-51

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