Abstract

An adenovirus (Ad) early region 3 (E3) protein called Ad E3-14.7K, which inhibits tumor necrosis factor alpha (TNFalpha) cytolysis, has been used in the yeast two hybrid system to isolate interacting proteins from human cells. The studies were initiated in order to determine the mechanism of action of Ad E3-14.7K. Four proteins called FIPs were detected in a HeLa cell cDNA library. Several of the FIPs interact with RIP (Fas receptor interacting protein), which is an important mediator of cell death induced by TNFalpha and the Fas ligand. FlPI is a novel member of the low molecular weight GTPase superfamily and also has a domain that is homologous to several bacterial proteases. FlPI also associates with a series of cell phosphoproteins following stimulation by TNFalpha and has been used as """"""""bait"""""""" to isolate additional cell proteins called GIPs. Several of the GIPs also interact with RIP. Each of the FIPs colocalizes with Ad E3- 14.7K in discrete foci in the cytoplasm. Further studies are proposed to: (1) determine how these various proteins interact physically and functionally in multiprotein complexes. This will be achieved by selected mutagenesis; (2) identify and determine the function of the phosphoproteins that interact with FlP1 and (3) determine if the FlP-1 homologue (ceT24fl) gene in c. elegans affects development. Control of cell death plays an important part in embryonic and fetal development, autoimmunity and cancer as well as other important human diseases. What we learn from studying viral genes that have evolved to prevent cell death by interacting with novel cellular targets should extend our understanding about a variety of human diseases affected by this basic process.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA072963-05
Application #
6376341
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Wong, May
Project Start
1997-07-23
Project End
2004-04-30
Budget Start
2001-05-01
Budget End
2004-04-30
Support Year
5
Fiscal Year
2001
Total Cost
$311,948
Indirect Cost

  Institution

Name
Albert Einstein College of Medicine
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461

  Publications

Shifera, Amde Selassie; Horwitz, Marshall S (2008) Mutations in the zinc finger domain of IKK gamma block the activation of NF-kappa B and the induction of IL-2 in stimulated T lymphocytes. Mol Immunol 45:1633-45
Shifera, Amde Selassie; Friedman, Joshua M; Horwitz, Marshall S (2008) IKK gamma (NEMO) is involved in the coordination of the AP-1 and NF-kappa B pathways. Mol Cell Biochem 310:181-90
Horwitz, Marshall S (2004) Function of adenovirus E3 proteins and their interactions with immunoregulatory cell proteins. J Gene Med 6 Suppl 1:S172-83
Lesokhin, Alexander M; Delgado-Lopez, Fernando; Horwitz, Marshall S (2002) Inhibition of chemokine expression by adenovirus early region three (E3) genes. J Virol 76:8236-43
Friedman, Joshua M; Horwitz, Marshall S (2002) Inhibition of tumor necrosis factor alpha-induced NF-kappa B activation by the adenovirus E3-10.4/14.5K complex. J Virol 76:5515-21
Horwitz, M S (2001) Adenovirus immunoregulatory genes and their cellular targets. Virology 279:1-8
Tarassishin, L; Horwitz, M S (2001) Sites on FIP-3 (NEMO/IKKgamma) essential for its phosphorylation and NF-kappaB modulating activity. Biochem Biophys Res Commun 285:555-60
Ye, J; Xie, X; Tarassishin, L et al. (2000) Regulation of the NF-kappaB activation pathway by isolated domains of FIP3/IKKgamma, a component of the IkappaB-alpha kinase complex. J Biol Chem 275:9882-9
Lukashok, S A; Tarassishin, L; Li, Y et al. (2000) An adenovirus inhibitor of tumor necrosis factor alpha-induced apoptosis complexes with dynein and a small GTPase. J Virol 74:4705-9
Li, Y; Kang, J; Friedman, J et al. (1999) Identification of a cell protein (FIP-3) as a modulator of NF-kappaB activity and as a target of an adenovirus inhibitor of tumor necrosis factor alpha-induced apoptosis. Proc Natl Acad Sci U S A 96:1042-7

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