A series of mutants of the ME-180 (human cervical carcinoma) cell line resistant to the cytotoxic effects of interferon-gamma have been isolated. Such mutants have been found to be defective in indoleamine 2,3-dioxygenase (IDO) induction, the first enzyme of the tryptophan catabolic pathway. Second round mutants have also been isolated lacking completely IDO induction and affected also in other interferon-inducible responses, such as HLA and oligo A synthetase induction. The relationship between IDO induction, tryptophan transport and metabolism will be investigated in order to ascertain the physiological significance of this pathway. Using a cloned cDNA we shall investigate whether the IDO-mutants are point mutations or result from mutation in regulatory sequences using gel- retardation assays. Northern blot and Southern analyses will be done with """"""""global"""""""" mutants to investigate whether such mutants result from deletions or alterations in the signalling pathway. Receptor binding studies using I125 interferon will also be done. The structure of interferon responsive sequences in IDO genomic DNA will be investigated. The regulation of this gene by interferon-gamma will be investigated using mutant and wild-type cells. Finally a genetic analysis of the mutants will be undertaken to locate the gene(s) affected to their respective chromosomes, and to ascertain whether the mutations are allelic.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA053783-03
Application #
2095482
Study Section
Medical Biochemistry Study Section (MEDB)
Project Start
1991-02-01
Project End
1994-01-31
Budget Start
1993-02-01
Budget End
1994-01-31
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Indiana University Bloomington
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
Townsend, Danyelle M; Findlay, Victoria J; Fazilev, Farit et al. (2006) A glutathione S-transferase pi-activated prodrug causes kinase activation concurrent with S-glutathionylation of proteins. Mol Pharmacol 69:501-8
McIlwain, C C; Townsend, D M; Tew, K D (2006) Glutathione S-transferase polymorphisms: cancer incidence and therapy. Oncogene 25:1639-48
Klein, S B; Yeivin, A; Becker, G et al. (1994) IDO mutants cross resistant to type I interferon retain p91-dependent gene induction. J Interferon Res 14:333-41
Ozes, O N; Taylor, M W (1994) Reversal of interferon-gamma-resistant phenotype by poly(I:C): possible involvement of ISGF2 (IRF1) in interferon-gamma-mediated induction of the IDO gene. J Interferon Res 14:25-32
Reiter, Z; Tomson, S; Ozes, O N et al. (1993) Combination treatment of 2-chlorodeoxyadenosine and type I interferon on hairy cell leukemia-like cells: cytotoxic effect and MHC-unrestricted killer cell regulation. Blood 81:1699-708
Burkin, D J; Kimbro, K S; Barr, B L et al. (1993) Localization of the human indoleamine 2,3-dioxygenase (IDO) gene to the pericentromeric region of human chromosome 8. Genomics 17:262-3
Reiter, Z; Taylor, M W (1993) Interleukin 2 protects hairy leukemic cells from lymphokine-activated killer cell-mediated cytotoxicity. Cancer Res 53:3555-60
Ozes, O N; Taylor, M W (1993) Reversal of an interferon-gamma-resistant phenotype by poly(I:C): possible role of double-stranded RNA-activated kinase in interferon-gamma signaling. J Interferon Res 13:283-8
Reiter, Z; Ozes, O N; Blatt, L M et al. (1992) Cytokine and natural killing regulation of growth of a hairy cell leukemia-like cell line: the role of interferon-alpha and interleukin-2. J Immunother (1991) 11:40-9
Reiter, Z; Ozes, O N; Blatt, L M et al. (1992) A dual anti-tumor effect of a combination of interferon-alpha or interleukin-2 and 5-fluorouracil on natural killer (NK) cell-mediated cytotoxicity. Clin Immunol Immunopathol 62:103-11

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