Signal Transduction and Activation of Gene Expression by Cytokines & Cellular Defense Responses to Viral Infection

Ph.D., 1983, State University of New York at Stony Brook
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Cytokines

Cells respond to their environment with rapid and specific biological changes that can lead to proliferation, growth inhibition, differentiation, or cellular suicide. Polypeptide hormones (cytokines) can elicit these rapid physiological responses by binding to cell surface receptors and subsequently transmitting a signal to the nucleus that activates the transcription of a specific set of genes.

The signal transduction pathway of one class of cytokines initiates with the activation of tyrosine kinases of the Janus kinase (JAK) family. JAKs are associated with the cytoplasmic domains of cytokine transmembrane receptors. Following activation they phosphorylate the receptor and latent cytoplasmic transcription factors. The transcription factors are members of the family of signal transducers and activators of transcription (STATs). Tyrosine phosphorylation of the STATs promotes the formation of homomeric or heteromeric protein complexes that translocate to the nucleus, bind to specific DNA target sites in the promoters of induced genes, and stimulate gene transcription.

Many cytokines that use a JAK/STAT signal pathway function to enhance native and acquired immunity. Our ongoing studies with interferons, interleukin-2, interleukin-4, interleukin-6, and prolactin characterize the diverse physiological effects of cytokines, the molecular interactions that specify distinct STAT activation, the cytoplasmic/ nuclear localization of the STATs, the effect of oncoproteins on the STATs, and the role of JAK/STAT pathway in Drosophila development.

Viral Defense

Mammalian cells also respond to viral infection with the expression of new genes that function in host survival. During viral infection with DNA or RNA viruses, viral dsRNA is generated by transcription and/or replication. This dsRNA is a potent intracellular signal that stimulates the defense response of the cell. One of the signal transduction pathways activated by dsRNA leads to the transcriptional induction of type I interferon genes. Interferons are cytokines that have the unique abilty to confer resistance to viral infections, inhibit the growth of normal and tumorigenic cells, and induce the differentiation and activation of a variety of immune cells. The initial cell that is infected and producing interferon may gain little benefit from its action. Our investigations have led to the discovery of another primary defense response of the cell that is stimulated by dsRNA and is independent of interferon. DsRNA activates a latent transcription factor, designated the dsRNA activated factor 1 (DRAF1) that directly induces a subset of interferon stimulated genes. Two components of DRAF1 have been identified:Interferon Regulatory Factor -3 (IRF-3), and the histone acetylases, CREB binding protein (CBP) and p300. IRF-3 resides in the cytoplasm until it is specifically phosphorylated and then it translocates to the nucleus.

Current studies on the DRAF1 defense pathway include identification of the kinase that is activated by viral dsRNA to phosphorylate IRF-3, characterization of mechanisms that control nuclear-cytoplasmic localization, identification of genes that are activated specifically by DRAF1,and determination of the physiological effects of DRAF1 activation.

Selected Publications

• McBride Kevin M; Banninger Gregg; McDonald Christine; Reich Nancy C. Regulated nuclear import of the STAT1 transcription factor by direct binding of importin-alpha. EMBO (European Molecular Biology Organization) Journal. 21. April 2, 2002. 1754-1763.
  
  
• Faruqi. T.R., D. Gomez, X. Bustelo, D. Bar-Sagi and N.C. Reich. 2001. Rac1 mediates STAT3 activation by autocrine IL-6. Proc Natl Acad Sci USA, 98:9014-9019
  
  
• Weaver, B., O. Ando, K.P. Kumar and N.C. Reich. 2001. Apoptosis promoted by the dsRNA activated factor (DRAF1) during viral infection independent of the action of interferon or p53. FASEB J., 15:501-551
  
  
• McBride, K.E., C. McDonald, and N.C. Reich. 2000. Nuclear export signal located within the DNA-binding domain of the STAT1 transcription factor. EMBO J. 19:6196-6206
  
  
• Kumar, K.P., K. McBride, B. Weaver, C. Dingwall, and N.C. Reich. 2000. Regulated nuclear-cytoplasmic localization of interferon regulatory factor-3 subunit of the dsRNA activated transcription factor, DRAF1. Mol. & Cell. Biol. 20:4159-4168.
  
  
• McDonald, C. and N.C. Reich. 1999. Cooperation of the transcriptional coactivators CBP/p300 with STAT6. J. Interferon & Cytokine Res. 19:711-722
  
  
• Weaver, B.K., K.P. Kumar and N.C. Reich. 1998. Interferon Regulatory Factor 3 and CREB-Binding Protein/p300 are Subunits of the dsRNA Activated Transcription Factor, DRAF1. Mol. & Cell. Biology 18:359-368.
  
  
• Martinez-Moczygemba, M., M.J. Gutch, D. French and N.C. Reich. 1997. Distinct STAT Structure Promotes Interaction of STAT2 with the p48 Subunit of the Interferon-a Stimulated Transcription Factor, ISGF3. J. Biol. Chem. 272:200070-200076.