Airway inflammatory diseases continue to inflict significant morbidity in the US. NF-KB/RelA is a cytosolic transcription factor that mediates the inflammatory response to cytokines, viruses, oxidative stressors, and allergens. Our studies in the past project period have demonstrated that the NF-KB/RelA activation pathway is bipartite, with one arm releasing RelA from cytoplasmic inhibition, and a separate arm inducing reactive oxygen species (ROS)-dependent, site-specific RelA Ser[276] phosphorylation, required for activating inflammation. Our preliminary studies using TNF-a as stimulus have shown that RelA Ser[276] phosphorylation is a switch required for RelA association with a complex containing the cyclin-dependent kinase (CDK)-9 and bromodomain-4 (Brd4) proteins. CDK9 phosphorylates the C-terminus of RNA Polymerase II, promoting entry into transcriptional elongation mode that produces full-length mRNA transcripts. During this next grant period P1 seeks to understand the mechanism(s) by which RSV modulates NF-KB signaling in the epithelial cell. Our central hypothesis is that NF-KB /RelA Ser[276] phosphorylation couples to an acetyl (Ac) RelA- Brd4-CDK9 pathway that mediates transcriptional elongation as a mechanism for inflammatory gene expression, leading to airway inflammation.
Aim 1 will test the hypothesis that RelA Ser[310] phosphorylation is required for RelA Lys[310] acetylation and Brd4-CDK9 recruitment to promote inflammatory gene expression in response to RSV and 8-oxoG in cultured airway cells.
Aim 2 will test the hypothesis that CDK9 mediates RNA Pol II C-terminal domain (CTD) phosphorylation, clearance and transcriptional elongation in NF-KB-dependent gene expression in cultured airway cells.
Aim 3 will test the hypothesis that NF-KB-induced CDK9 recruitment mediates transcriptional elongation in RSV-induced mucosal inflammation in a mouse model in vivo. This project is highly synergistic with Projects 2-4, informing hypothesis testing and shared experimental work, and relies upon the support of Cores A and B. PI is innovative because it will for the first time establish the mechanism for the RelA phosphorylation/acetylation-transcriptional elongation pathway in RSV and 8-oxoGinduced inflammatory gene expression, and establish this pathway as a central mediator of airway inflammation. These discoveries will lay the foundation for new approaches to selective NF-KB inhibition, for example using CDK9 inhibitors.

Public Health Relevance

Asthma is a breathing disorder affecting about 10% of Americans, and is frequently worsened by cold viruses. This project seeks to understand how the lung tissues respond to signals produced by cold viruses that worsen the disease. This knowledge should lead to novel approaches to asthma treatment, based on blocking the signals elucidated in this project.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
2P01AI062885-06A1
Application #
8465464
Study Section
Allergy & Clinical Immunology-1 (AITC)
Project Start
2005-07-01
Project End
2017-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
6
Fiscal Year
2012
Total Cost
$297,733
Indirect Cost
$102,656
Name
University of Texas Medical Br Galveston
Department
Type
DUNS #
800771149
City
Galveston
State
TX
Country
United States
Zip Code
77555
Bazhanov, Nikolay; Escaffre, Olivier; Freiberg, Alexander N et al. (2017) Broad-Range Antiviral Activity of Hydrogen Sulfide Against Highly Pathogenic RNA Viruses. Sci Rep 7:41029
Tian, Bing; Patrikeev, Igor; Ochoa, Lorenzo et al. (2017) NF-?B Mediates Mesenchymal Transition, Remodeling, and Pulmonary Fibrosis in Response to Chronic Inflammation by Viral RNA Patterns. Am J Respir Cell Mol Biol 56:506-520
Zhang, Yueqing; Sun, Hong; Zhang, Jing et al. (2017) Quantitative Assessment of the Effects of Trypsin Digestion Methods on Affinity Purification-Mass Spectrometry-based Protein-Protein Interaction Analysis. J Proteome Res 16:3068-3082
Agod, Zsofia; Fekete, T√ľnde; Budai, Marietta M et al. (2017) Regulation of type I interferon responses by mitochondria-derived reactive oxygen species in plasmacytoid dendritic cells. Redox Biol 13:633-645
Zhao, Yingxin; Jamaluddin, Mohammad; Zhang, Yueqing et al. (2017) Systematic Analysis of Cell-Type Differences in the Epithelial Secretome Reveals Insights into the Pathogenesis of Respiratory Syncytial Virus-Induced Lower Respiratory Tract Infections. J Immunol 198:3345-3364
Komaravelli, Narayana; Ansar, Maria; Garofalo, Roberto P et al. (2017) Respiratory syncytial virus induces NRF2 degradation through a promyelocytic leukemia protein - ring finger protein 4 dependent pathway. Free Radic Biol Med 113:494-504
German, Peter; Saenz, David; Szaniszlo, Peter et al. (2017) 8-Oxoguanine DNA glycosylase1-driven DNA repair-A paradoxical role in lung aging. Mech Ageing Dev 161:51-65
Tian, Bing; Yang, Jun; Zhao, Yingxin et al. (2017) BRD4 Couples NF-?B/RelA with Airway Inflammation and the IRF-RIG-I Amplification Loop in Respiratory Syncytial Virus Infection. J Virol 91:
Ke, Yueshuang; Han, Yanlong; Guo, Xiaolan et al. (2017) PARP1 promotes gene expression at the post-transcriptiona level by modulating the RNA-binding protein HuR. Nat Commun 8:14632
Bazhanov, Nikolay; Ansar, Maria; Ivanciuc, Teodora et al. (2017) Hydrogen Sulfide: A Novel Player in Airway Development, Pathophysiology of Respiratory Diseases, and Antiviral Defenses. Am J Respir Cell Mol Biol 57:403-410

Showing the most recent 10 out of 145 publications