The COP9 signalosome (CSN) complex is an evolutionally conserved protein complex present in all eukaryotes and is involved in controlling diverse cellular and developmental processes. Disregulation of the CSN complex can have a dramatic effect on cellular functions critical to tumor development, including maintenance of DNA fidelity, cell cycle control, DNA repair, angiogenesis, and micro-environmental homeostasis. The CSN complex has recently been found to be involved in the regulation of NF-?B activated by excellular signaling. But a full understanding of the function and regulation of the CSN complex in signal-induced NF-?B activation pathways is lacking. NF-?B proteins are inducible transcription factors that are crucial regulators of many physiological and pathophysiological processes. Aberrant regulation of NF-?B and the signaling pathways (e.g. TNFa pathway) controlling its activity are involved in cancer development and progression, as well as resistance to chemotherapy and radiotherapy. Therefore, NF-?B and components involved in regulating its activity have become a focal point for drug discovery. Given its critica importance in cancer development, we hypothesize that detailed analysis of the CSN complex in the NF-?B pathway will not only provide new insights into its function and regulation in general, but also lead to new directions in future development of clinically useful inhibitors for cancer treatment targeting the NF-?B system through the CSN complex. To test this, we propose to achieve the following specific aims: 1) To define protein interaction dynamics of the human CSN complex associated with NF-?B activation triggered by TNFa signaling. 2) To characterize CSN5 phosphorylation and the dynamic interactions of the CSN5-IKK complex upon TNFa signaling. The proposed work represents the first detailed proteomic analysis of the CSN complex in the context of signaling pathways. The methodology developed in this work will be applicable for the study of other protein complexes in response to extracellular signaling.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Exploratory/Developmental Grants (R21)
Project #
Application #
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Knowlton, John R
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of California Irvine
Anatomy/Cell Biology
Schools of Arts and Sciences
United States
Zip Code
Burke, Anthony M; Kandur, Wynne; Novitsky, Eric J et al. (2015) Synthesis of two new enrichable and MS-cleavable cross-linkers to define protein-protein interactions by mass spectrometry. Org Biomol Chem 13:5030-7
Kandur, Wynne V; Kao, Athit; Vellucci, Danielle et al. (2015) Design of CID-cleavable protein cross-linkers: identical mass modifications for simpler sequence analysis. Org Biomol Chem 13:9793-807
Li, Qiuling; Li, Yuewei; Gu, Bingnan et al. (2015) Akt Phosphorylates Wnt Coactivator and Chromatin Effector Pygo2 at Serine 48 to Antagonize Its Ubiquitin/Proteasome-mediated Degradation. J Biol Chem 290:21553-67
Zhang, Hong-Hai; Wang, Wen; Feng, Lin et al. (2015) S-nitrosylation of Cofilin-1 Serves as a Novel Pathway for VEGF-Stimulated Endothelial Cell Migration. J Cell Physiol 230:406-17
Chong, Robert A; Wu, Kenneth; Spratt, Donald E et al. (2014) Pivotal role for the ubiquitin Y59-E51 loop in lysine 48 polyubiquitination. Proc Natl Acad Sci U S A 111:8434-9
Yu, Clinton; Kandur, Wynne; Kao, Athit et al. (2014) Developing new isotope-coded mass spectrometry-cleavable cross-linkers for elucidating protein structures. Anal Chem 86:2099-106
Aphasizheva, Inna; Zhang, Liye; Wang, Xiaorong et al. (2014) RNA binding and core complexes constitute the U-insertion/deletion editosome. Mol Cell Biol 34:4329-42
Wang, Xiaorong; Huang, Lan (2014) Defining dynamic protein interactions using SILAC-based quantitative mass spectrometry. Methods Mol Biol 1188:191-205
Kaake, Robyn M; Wang, Xiaorong; Burke, Anthony et al. (2014) A new in vivo cross-linking mass spectrometry platform to define protein-protein interactions in living cells. Mol Cell Proteomics 13:3533-43
Kaake, Robyn M; Kao, Athit; Yu, Clinton et al. (2014) Characterizing the dynamics of proteasome complexes by proteomics approaches. Antioxid Redox Signal 21:2444-56