Project 1 (Brasier, A.R.)
Allan R. Brasier MD
Nelda C. and H.J. Lutcher Stark
Distinguished Professor in Internal Medicine
Department of Internal Medicine-Endocrinology
Sealy Center for Molecular Sciences
301 University Blvd.
Galveston, TX 77555-1060
Dr. Brasier's bio...
Asthma is a chronic inflammatory disorder of the airways, where it is known that viral infections play an important role in the manifestations of this disease. In particular, respriatory syncytial virus(RSV) is a common cause of episodic wheezing in children. Our work has shown that the airway epithelium responds to cytokines and viruses through changes in its genetic program, an important event in the initiation of pulmonary inflammation. We have shown that cytokine (TNF) stimulation and RSV replication both activate the NF-kB transcription factor that alters the expression of specific inflammatory genetic programs. In collaboration with Dr. Garofalo's group, we have also found that inhibiting NF-kB activity reduces lung inflammation in a mouse model of RSV induced inflammation. For this reason identification of the genes controlled by NF-kB have been an important focus of our research. Using high density oligonucleotide microarrays, we have identified groups of genes downstream of NF-kB in response to RSV and the inflammatory cytokine, TNF.
In this project, we are investigating our recent novel experimental findings that NF-kB-dependent genes appear to be activated in 3 distinct kinetic groups. We have also identified a reactive oxygen species (ROS)- sensitive step important in control of NF-kB dependent genes, where target gene expression can be inhibited after NF-kB binds DNA, and have shown that NF-kB/Rel A forms a nuclear complex with the NF-kB inducing kinase (NIK), a MAP3K kinase that forms a DNA- associated complex with IkB Kinase-a (IKKa) and the p300 coactivator.
We are pursuing the hypothesis that the NF-kB transcription factor is a central regulator of airway inflammation in the stimulated epithelial cell. NF-kB activation of target promoters requires chromatin remodeling, is redox-sensitive, and requires formation of a macromolecular complex with NIK, IKKa and p300. Our aims are to:
- Identify mechanisms for distinct temporal genomic responses to NF-kB using Chromatin immunoprecipitation (ChIP) assays.
- Map interacting domains of the NF-kB inducing kinase (NIK) –with NF-kB/Rel A and the p300 coactivator, determine their functional consequences and explore NIK’s role in the epithelial NF-kB dependent gene network.
- Determine the role of the NIK-IKKa , NIK-Rel A, and NIK- p300 complexes in NF-kB dependent gene expression.
- Develop a protein interaction network map of NF-kB/Rel A, IKKa, and NIK in cytokine stimulated and viral infected epithelial cells.
These studies will identify key intracellular signaling events in the NF-kB pathway controlling airway inflammation.
- Tian, B., Zhang, Y., Luxon, B.A., Garofalo, R.P., Casola, A., Sinha, M., and Brasier, A.R. † Identification Of NF-kB Dependent Gene Networks In Respiratory Syncytial Virus-Infected Cells. Journal of Virology, 2002; 76: 6800-6814.
- Haeberle, H., Takizawa, R., Casola, A., Brasier, A.R., Hans-Juergen D., van Rooijen, R., Gatalica, Z., Garofalo, R.P. Respiratory syncytial virus-induced activation of NF-kB in the lung involves alveolar macrophages and Toll-like receptor 4-dependent pathways. J Infect Dis 2002; 186:1199-1206.
- Zhang,, Y., Jamaluddin, M., Tian, B., Wang, S., Casola A. Garofalo, R., and Brasier, AR. Ribavirin Treatment Upregulates Anti-Viral Gene Expression Via The IFN-Stimulated Response Element In RSV-Infected Epithelial Cells. Journal of Virology, 2003; 78: 5933-5947.
- Page, K.; Zhou, L.; Iazvovskaia, S.;Corbit, K.C.;Soh, J.-W.; Weinstein, I.B.; Brasier, A.R.; and Hershenson, M.B. Regulation of airway epithelial cell NF-kB dependent gene expression by protein kinase Cd. Journal of Immunology, 2003; 170: 5681-9.
- Liu, T., Castro, S., Jamaluddin, M., Brasier, A.R., Garofalo, R.P. and Casola, A. ROS mediate viral-induced STAT activation: role of tyrosine phosphatases. J. Biol Chem, 2004; 279: 2461-2469.
- Haeberle, H., Casola, A., Gatalica, Z., Petronella S., Dieterich, H.-J., Ernst, P.B., Brasier, A.R., Garofalo, R.P. IkB kinase is a critical regulator of chemokine expression and lung inflammation in respiratory syncytial virus infection. Journal of Virology, 2004; 78: 2232-2241.
- Lipniaki, T., Paszek, P., Brasier, A.R., Luxon, B. and Kimmel, M. Mathematical model of NF-kB regulatory module. Journal of Theoretical Biology, 2004; 228: 195-215.
- Yamaoka, Y., Kudo, T., Lu, H., Casola, A., Brasier, A.R., Graham, D.Y. Role of Interferon Stimulated Response-Like Element in IL-8 promoter in Helicobacter pylori infection. Gastroenterology, 2004; 126: 1030-1043.
- Brasier, AR, Spratt, H. Wu, Z., Boldogh, S., Zhang, Y., Garofalo, R.P., Casola, A., Pashmi, J., Haag, A., Luxon, B., and Kurosky, A. Nuclear Heat Shock Response and Novel Nuclear Domain 10 Reorganization in Respiratory Syncytial Virus-Infected A549 Cells Identified By High Resolution 2D Gel Electrophoresis. Journal of Virology, 2004; 78: 11461-11476.
- Tian, B., Nowak, D., Jamaluddin, M., Wang, S. and Brasier, A.R. Identification Of Direct Genomic Targets Downstream Of The NF-kB Transcription Factor Mediating TNF Signaling. Journal of Biological Chemistry, 2005; 280:17435-17448.
- Tian, B., Nowak, D., and Brasier, A.R A TNF Induced Gene Expression Program Under Oscillatory NF-kB Control. BMC Genomics, 6:137, 2005.
- Jamaluddin, M. Choudhary, S., Wang, S., Casola, A., Huda, R., Garofalo,R.P., Ray, S. and Brasier, A.R. RSV-Inducible Bcl-3 Expression Antagonizes The Stat/IRF And NF-kB Signaling Pathways By Inducing HDAC-1 Recruitment To The IL-8 Promoter. Journal of Virology,2006; 79(24),15302-15313.