The physiological state of a cell is modulated in response to specific extracellular signals through changes in metabolic processes resulting, in part, from differential regulation of gene expression. The broad objective of this proposal is to characterize the molecular mechanisms involved in signal transduction initiated by polypeptide ligands binding to their specific cell surface receptors. Interferons (IFNs) area a class of polypeptide cytokines which induce rapid and coordinate transcriptional activation of a set of dispersed interferon stimulated genes (ISGs), leading to profound effects on the growth, differentiation, and metabolism of sensitive cells. The activation of these genes is completely dependent on the presence of IFN bound to its cell surface receptor and is mediated through the action of pre-existing protein components which change in activity, rather than in abundance, following treatment with IFN. Although the cis-acting DNA sequences regulating these genes have been characterized and trans-acting nuclear and cytoplasmic factors have been identified, the protein components of the signal transduction pathway from the cell surface to the nucleus, the nature of the biochemical events which carry the signal, and the mechanisms of transcriptional control of ISGs remain at present largely undefined. The major goal of this project is to develop a comprehensive understanding of IFN-induced transcription which leads to production of gene products necessary for growth control and for viral resistance. We propose to determine the molecular mechanics of IFN-induced signal transduction by defining genes and gene products in the pathway, by purifying and characterizing constituent proteins, and by defining the biochemical steps involved in signal generation. In this proposal, the individual components of signal transduction will be isolated and purified in order to reconstitute the process in vitro and to study the biochemical steps involved. This will be approached by isolating, purifying, and cloning the positive transcription factor which activates ISG expression in the nucleus, the protein precursors to this transcription factor which exists in the cytoplasm, the proteins which interact with these components to achieve activation and nuclear translocation, the type I IFN cell surface receptor which initially generates the signal, and membrane and cytoplasmic receptor-proximal proteins which serve as links in the signal chain. These proteins will be analyzed in vitro and in vivo by mutagenesis to define functional domains involved in IFN binding, signal transduction, subunit association, nuclear translocation, DNA binding, and transcriptional activation. Knowledge of the normal mechanism of action of IFN will provide a basis for understanding its role in viral inhibition and control of proliferation and will lead to investigations of defects in these processes which may contribute to impaired resistance to infectious disease and to abnormal cell growth. These studies may also be broadly applicable signal transduction systems involving polypeptide hormones and growth factors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI028900-01
Application #
3143551
Study Section
Virology Study Section (VR)
Project Start
1990-01-01
Project End
1994-12-31
Budget Start
1990-01-01
Budget End
1990-12-31
Support Year
1
Fiscal Year
1990
Total Cost
Indirect Cost
Name
New York University
Department
Type
Schools of Medicine
DUNS #
004514360
City
New York
State
NY
Country
United States
Zip Code
10012
Genini, Davide; Brambilla, Lara; Laurini, Erik et al. (2017) Mitochondrial dysfunction induced by a SH2 domain-targeting STAT3 inhibitor leads to metabolic synthetic lethality in cancer cells. Proc Natl Acad Sci U S A 114:E4924-E4933
Garama, Daniel J; Harris, Tiffany J; White, Christine L et al. (2015) A Synthetic Lethal Interaction between Glutathione Synthesis and Mitochondrial Reactive Oxygen Species Provides a Tumor-Specific Vulnerability Dependent on STAT3. Mol Cell Biol 35:3646-56
Gough, Daniel J; MariƩ, Isabelle J; Lobry, Camille et al. (2014) STAT3 supports experimental K-RasG12D-induced murine myeloproliferative neoplasms dependent on serine phosphorylation. Blood 124:2252-61
Gnatovskiy, Leonid; Mita, Paolo; Levy, David E (2013) The human RVB complex is required for efficient transcription of type I interferon-stimulated genes. Mol Cell Biol 33:3817-25
Gough, Daniel J; Koetz, Lisa; Levy, David E (2013) The MEK-ERK pathway is necessary for serine phosphorylation of mitochondrial STAT3 and Ras-mediated transformation. PLoS One 8:e83395
Gough, Daniel J; Messina, Nicole L; Clarke, Christopher J P et al. (2012) Constitutive type I interferon modulates homeostatic balance through tonic signaling. Immunity 36:166-74
Levy, David E; MariƩ, Isabelle J (2012) STATus report on tetramers. Immunity 36:553-5
Lee, Jason E; Yang, Yang-Ming; Liang, Feng-Xia et al. (2012) Nongenomic STAT5-dependent effects on Golgi apparatus and endoplasmic reticulum structure and function. Am J Physiol Cell Physiol 302:C804-20
Zhang, Liang; Das, Priyabrata; Schmolke, Mirco et al. (2012) Inhibition of pyrimidine synthesis reverses viral virulence factor-mediated block of mRNA nuclear export. J Cell Biol 196:315-26
Corcoran, Ryan B; Contino, Gianmarco; Deshpande, Vikram et al. (2011) STAT3 plays a critical role in KRAS-induced pancreatic tumorigenesis. Cancer Res 71:5020-9

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