The long-term goal of this study is to understand the biological function of protein modification by the ubiquitin like protein ISG15. In recent years our knowledge of protein ubiquitination has expanded rapidly and as a consequence protein ubiquitination has been found to play important roles in various aspects of cellular functions, including cell cycle, membrane receptor signal transduction, endocytosis, and DNA repair. In contrast, little is known about the role of protein modification by ISG15. We cloned a novel member of the deubiquitinating enzyme family, termed UBP43. Further studies have demonstrated that UBP43 is a protease that specifically removes ISG15 from its conjugates. Type I interferon (INFa/b) and LPS strongly upregulate both UBP43 expression and protein ISGylation. We have generated UBP43 knockout mice. UBP43 deficient mice show fundamental defects in development and in response to interferon and LPS treatment. This proposal will test the hypothesis that protein ISG15 modification plays a fundamental role in maintaining cellular metabolism under conditions of stress. The studies proposed in Specific Aim#1 will identify the molecular targets of ISG15. We have established an immuno-affinity purification method for such an approach. The studies proposed in Specific Aim #2 will characterize the function of protein ISGylation by defining ISG15 modification sites in target proteins and studying the effect of ISGylation on target proteins. We have identified several ISGylated proteins. The studies proposed in Specific Aim #3 will generate conditional ISG15 activating enzyme UBE1L knockout mice to analyze the function of protein ISGylation. The experiments proposed will address important questions about protein ISGylation and may provide valuable insights into the therapeutic treatment of human diseases, such as viral infection and cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM066955-04
Application #
7085433
Study Section
Biochemistry Study Section (BIO)
Program Officer
Jones, Warren
Project Start
2003-07-01
Project End
2008-02-29
Budget Start
2006-07-01
Budget End
2008-02-29
Support Year
4
Fiscal Year
2006
Total Cost
$333,847
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Zhang, Dongxian; Zhang, Dong-Er (2011) Interferon-stimulated gene 15 and the protein ISGylation system. J Interferon Cytokine Res 31:119-30
Nakka, Venkata P; Lang, Bradley T; Lenschow, Deborah J et al. (2011) Increased cerebral protein ISGylation after focal ischemia is neuroprotective. J Cereb Blood Flow Metab 31:2375-84
Maragno, Ana Leticia; Pironin, Martine; Alcalde, Helene et al. (2011) ISG15 modulates development of the erythroid lineage. PLoS One 6:e26068
Kawabe, Hiroshi; Neeb, Antje; Dimova, Kalina et al. (2010) Regulation of Rap2A by the ubiquitin ligase Nedd4-1 controls neurite development. Neuron 65:358-72
Cong, Xiuli; Yan, Ming; Yin, Xiaoyan et al. (2010) Hematopoietic cells from Ube1L-deficient mice exhibit an impaired proliferation defect under the stress of bone marrow transplantation. Blood Cells Mol Dis 45:103-11
Lai, Caroline; Struckhoff, Jessica J; Schneider, Jana et al. (2009) Mice lacking the ISG15 E1 enzyme UbE1L demonstrate increased susceptibility to both mouse-adapted and non-mouse-adapted influenza B virus infection. J Virol 83:1147-51
Yin, Xiaoyan; Cong, Xiuli; Yan, Ming et al. (2009) Deficiency of a potential 3p21.3 tumor suppressor gene UBE1L (UBA7) does not accelerate lung cancer development in K-rasLA2 mice. Lung Cancer 63:194-200
Yin, Xiaoyan; Cong, Xiuli; Yan, Ming et al. (2009) Alteration of tumor spectrum by ISGylation in p53-deficient mice. Cancer Biol Ther 8:1167-72
Okumura, Fumihiko; Lenschow, Deborah J; Zhang, Dong-Er (2008) Nitrosylation of ISG15 prevents the disulfide bond-mediated dimerization of ISG15 and contributes to effective ISGylation. J Biol Chem 283:24484-8
Malakhova, Oxana A; Zhang, Dong-Er (2008) ISG15 inhibits Nedd4 ubiquitin E3 activity and enhances the innate antiviral response. J Biol Chem 283:8783-7

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