Mitogen-activated protein kinase (MARK) pathways are evolutionary-conserved eukaryotic signaling modules that regulate diverse cellular processes. In response to a multitude of extracellular stimuli, MAPKs phosphorylate and activate downstream transcription factors that modify chromatin and orchestrate gene expression. Many MARK pathways have been genetically defined in Saccharomyces cerevisiae. In this proposal we focus on one of the well-characterized yeast MARK pathways, the high osmolarity glycerol (HOG) response pathway. During the last few decades, the components and the regulatory network of this cascade have been elucidated using genetic and biochemical assays performed on large populations of cells. In this proposal we take a complementary approach by monitoring the signaling dynamics in single cells with high temporal resolution. This method provides three distinct advantages compared to traditional biochemical and genetic assays on cell populations: (1) MARK activity will be measured in live cells;(2) measurements are almost instantaneous giving a temporal resolution on the order of 1 second which is the typical time-scale of the kinetics of individual reaction steps;(3) measurements will be performed on many single cells in parallel, allowing studies of cell-to-cell variability in signaling dynamics and allowing a direct measurement of the correlation between the concentration of a key component in the network and the signaling dynamics in a single cell.
The specific aims are organized around measuring the signaling dynamics at three different time-scales: (1) the initial rapid signal propagation before feedback regulation is activated;(2) adaptation dynamics dominated by feedback regulation and (3) evolutionary time-scales, which will be explored by using a comparative experimental analysis of the HOG signaling pathway of related yeast species. The proposed experimental work will be closely integrated with quantitative modeling approaches with the ultimate goal to build a predictive model of the HOG response pathway. Relevance to public health: Some mammalian MAPKs are activated by inflammatory cytokines and environmental stresses which might play an important role in diseases like asthma and autoimmunity. Because MARK cascades are highly conserved, the proposed experimental and theoretical techniques and concepts that will be important for the analysis of MARK cascades in higher eukaryotes.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
Modeling and Analysis of Biological Systems Study Section (MABS)
Program Officer
Brazhnik, Paul
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Massachusetts Institute of Technology
Schools of Arts and Sciences
United States
Zip Code
Soh, Y Q Shirleen; Junker, Jan Philipp; Gill, Mark E et al. (2015) A Gene Regulatory Program for Meiotic Prophase in the Fetal Ovary. PLoS Genet 11:e1005531
Slavov, Nikolai; Semrau, Stefan; Airoldi, Edoardo et al. (2015) Differential Stoichiometry among Core Ribosomal Proteins. Cell Rep 13:865-73
Slavov, Nikolai; Budnik, Bogdan A; Schwab, David et al. (2014) Constant growth rate can be supported by decreasing energy flux and increasing aerobic glycolysis. Cell Rep 7:705-14
Junker, Jan Philipp; Peterson, Kevin A; Nishi, Yuichi et al. (2014) A predictive model of bifunctional transcription factor signaling during embryonic tissue patterning. Dev Cell 31:448-60
Fang, Miaoqing; Xie, Huangming; Dougan, Stephanie K et al. (2013) Stochastic cytokine expression induces mixed T helper cell States. PLoS Biol 11:e1001618
Tan, Rui Zhen; Ji, Ni; Mentink, Remco A et al. (2013) Deconvolving the roles of Wnt ligands and receptors in sensing and amplification. Mol Syst Biol 9:631
Kim, Dong hyun; GrĂ¼n, Dominic; van Oudenaarden, Alexander (2013) Dampening of expression oscillations by synchronous regulation of a microRNA and its target. Nat Genet 45:1337-44
Teytelman, Leonid; Thurtle, Deborah M; Rine, Jasper et al. (2013) Highly expressed loci are vulnerable to misleading ChIP localization of multiple unrelated proteins. Proc Natl Acad Sci U S A 110:18602-7
Tan, Rui Zhen; van Oudenaarden, Alexander (2010) Transcript counting in single cells reveals dynamics of rDNA transcription. Mol Syst Biol 6:358
Dong, Guogang; Yang, Qiong; Wang, Qiang et al. (2010) Elevated ATPase activity of KaiC applies a circadian checkpoint on cell division in Synechococcus elongatus. Cell 140:529-39

Showing the most recent 10 out of 27 publications