Core B will provide proteomics and protein analysis services for PPG projects. Throughout the last funding cycle, the protein analysis component of Core B has significantly expanded to meet the needs of the individual projects in the PPG. In addition to mass spectrometry (MS)-based footprinting technology for the mapping of protein-protein and protein-nucleic acid interactions proposed in the previous application, we have added the following new services to the Core: 1) the AKTA-fast protein liquid chromatography (FPLC), which has played a critical role in obtaining purified recombinant proteins for biochemical/biophysical characterization for former Project 3 and antibody production for former Project 2;and 2) the EnSpire multimode plate reader to provide a new service for highly sensitive, rapid and robust monitoring of protein-protein and protein-nucleic acid interactions. MS-based proteomic approaches enabled us to identify new protein-protein interactions and posttranslational modifications, and have thus been instrumental for advancing the research of former Projects 1 and 2, respectively. The above technologies will continue to play an important role in accomplishing the research objectives of the PPG projects proposed in this competitive renewal. In particular, Specific Aims of the Proteomics and Protein Analysis Core are:
Aim 1 : to perform MS-based protein footprinting experiments to identify contact amino acids in the context of large biologically relevant nucleoprotein and protein-protein complexes;
Aim 2 : to develop novel applications of the EnSpire multimode plate reader for biochemical characterization of protein-nucleic acid and protein-protein interactions using homogeneous time-resolved fluorescence energy transfer (HTRF), AlphaLISA, and AlphaScreen (ALPHA) based technologies;
Aim 3 : to provide a column chromatography service using AKTA-FPLC to assist individual PPG projects to isolate low abundant native protein-protein complexes from mammalian cells and purify mg quantities of poorly soluble recombinant proteins from E. coli for their subsequent biochemical and biophysical characterization;
Aim 4 : to conduct MS-based proteomics experiments to identify interacting partners and protein posttranslational modifications. Taken together, Core B will provide the wide range of protein analysis services. Importantly, these services will be utilized by all four PPG projects.

Public Health Relevance

Overall goals of the PPG include delineating mechanisms of T-cell leukemia induced by the human T cell leukemia virus type 1 (HTLV-1). These studies require state-of-the-art proteomics and protein analysis technologies, which will be provided by Core B.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
2P01CA100730-11A1
Application #
8742037
Study Section
Special Emphasis Panel (ZCA1-RPRB-J (M1))
Project Start
2014-09-23
Project End
2019-08-31
Budget Start
2014-09-23
Budget End
2015-08-31
Support Year
11
Fiscal Year
2014
Total Cost
$176,110
Indirect Cost
$46,374
Name
Ohio State University
Department
Type
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Romeo, Megan; Hutchison, Tetiana; Malu, Aditi et al. (2018) The human T-cell leukemia virus type-1 p30II protein activates p53 and induces the TIGAR and suppresses oncogene-induced oxidative stress during viral carcinogenesis. Virology 518:103-115
Cherian, Mathew A; Olson, Sydney; Sundaramoorthi, Hemalatha et al. (2018) An activating mutation of interferon regulatory factor 4 (IRF4) in adult T-cell leukemia. J Biol Chem 293:6844-6858
Huey, Devra D; Bolon, Brad; La Perle, Krista M D et al. (2018) Role of Wild-type and Recombinant Human T-cell Leukemia Viruses in Lymphoproliferative Disease in Humanized NSG Mice. Comp Med 68:4-14
Pérès, Eléonore; Blin, Juliana; Ricci, Emiliano P et al. (2018) PDZ domain-binding motif of Tax sustains T-cell proliferation in HTLV-1-infected humanized mice. PLoS Pathog 14:e1006933
Panfil, Amanda R; Al-Saleem, Jacob; Howard, Cory M et al. (2018) Stability of the HTLV-1 Antisense-Derived Protein, HBZ, Is Regulated by the E3 Ubiquitin-Protein Ligase, UBR5. Front Microbiol 9:80
Murali, Bhavna; Ren, Qihao; Luo, Xianmin et al. (2018) Inhibition of the Stromal p38MAPK/MK2 Pathway Limits Breast Cancer Metastases and Chemotherapy-Induced Bone Loss. Cancer Res 78:5618-5630
Huey, Devra D; Niewiesk, Stefan (2018) Production of Humanized Mice through Stem Cell Transfer. Curr Protoc Mouse Biol 8:17-27
Kenney, Adam D; Dowdle, James A; Bozzacco, Leonia et al. (2017) Human Genetic Determinants of Viral Diseases. Annu Rev Genet 51:241-263
Webb, Lindsay M; Amici, Stephanie A; Jablonski, Kyle A et al. (2017) PRMT5-Selective Inhibitors Suppress Inflammatory T Cell Responses and Experimental Autoimmune Encephalomyelitis. J Immunol 198:1439-1451
Singh, Gatikrushna; Fritz, Sarah M; Ranji, Arnaz et al. (2017) Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution. J Vis Exp :

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