The overarching aim of this proposal is to understand the design principles of the regulatory network that regulates mitosis. The main focus is on the bistable trigger that governs the transition from interphase to mitosis. The proposed approaches include quantitative experimental studies in HeLa cells;Xenopus egg extracts, and reconstituted in vitro systems;fluorescent biosensor development;and computational studies. There are three specific aims: (1) To determine whether a bistable switch triggers the translocation of active cyclin B1/Cdk1 from the cytoplasm to the nucleus. Quantitative studies of the timing of early mitotic events in HeLa cells have led us to hypothesize that cyclin B1 is driven into the nucleus by a bistable circuit, where nuclear cyclin B1/Cdk1 complexes promote the import and/or inhibit the export of more cyclin B1/Cdk1 complexes. This hypothesis will be tested through experimental studies, and the implications of this hypothesis for noise propagation and self-organization will be explored through computational studies. (2) To determine how robust bistability is achieved by the bistable Cdk1/Cdc25/Wee1 system. Recent computational studies show that the striking 'mirror image'relationship between Cdc25 and Wee1 has the potential to render the bistability of the Cdk1/Cdc25/Wee1 nearly impervious to changes in protein concentration. We plan to test this hypothesis through reconstitution of the bistability switch in vitro and through perturbation of the system in extracto. (3) To develop a new class of fluorescent protein kinase sensors by exploiting the changes in bulk electrostatic properties that can occur when a protein is phosphorylated.

Public Health Relevance

The ability of a cell to replicate itself through growth, mitosis and cell division is one of the most basic, fundamental aspects of life. An understanding of the process also promises to shed light on cancer and cancer chemotherapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM061276-12
Application #
8197517
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Hamlet, Michelle R
Project Start
2000-04-01
Project End
2012-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
12
Fiscal Year
2012
Total Cost
$306,001
Indirect Cost
$114,881
Name
Stanford University
Department
Biology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Ferrell Jr, James E (2011) Simple rules for complex processes: new lessons from the budding yeast cell cycle. Mol Cell 43:497-500
Ferrell Jr, James E (2008) Feedback regulation of opposing enzymes generates robust, all-or-none bistable responses. Curr Biol 18:R244-5
Tsai, Tony Yu-Chen; Choi, Yoon Sup; Ma, Wenzhe et al. (2008) Robust, tunable biological oscillations from interlinked positive and negative feedback loops. Science 321:126-9
Hendrickson, David G; Hogan, Daniel J; Herschlag, Daniel et al. (2008) Systematic identification of mRNAs recruited to argonaute 2 by specific microRNAs and corresponding changes in transcript abundance. PLoS One 3:e2126
Santos, Silvia D M; Ferrell, James E (2008) Systems biology: On the cell cycle and its switches. Nature 454:288-9
Pomerening, Joseph R; Ubersax, Jeffrey A; Ferrell Jr, James E (2008) Rapid cycling and precocious termination of G1 phase in cells expressing CDK1AF. Mol Biol Cell 19:3426-41
Yue, Jianbo; Ferrell Jr, James E (2006) Mechanistic studies of the mitotic activation of Mos. Mol Cell Biol 26:5300-9
Pomerening, Joseph R; Kim, Sun Young; Ferrell Jr, James E (2005) Systems-level dissection of the cell-cycle oscillator: bypassing positive feedback produces damped oscillations. Cell 122:565-78
Yue, Jianbo; Ferrell Jr, James E (2004) Mos mediates the mitotic activation of p42 MAPK in Xenopus egg extracts. Curr Biol 14:1581-6
Angeli, David; Ferrell Jr, James E; Sontag, Eduardo D (2004) Detection of multistability, bifurcations, and hysteresis in a large class of biological positive-feedback systems. Proc Natl Acad Sci U S A 101:1822-7

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