Abstract

): The long-term objectives of this proposal are to develop further 5-aminoimidazole-4-carboxamide ribotide transformylase/inosine monophosphate cyclohydrolase (ATIC) as a c h e m otherapeutic target for the treatment of cancer and to create methotrexate-resistant variants of ATIC suitable for use in hematopoietic stem c e ll support thereby expanding the use of methotrexate in high-dose chemotherapy protocols.
The specific aims of this proposal are as follows: (1) to use pre-steady-state kinetic methods to define the minimal kinetic schemes for the two reactions catalyzed by ATIC; (2) to identify the regions of ATIC responsible for dimerization and substrate binding using approaches such as site-directed mutagenesis and protein footprinting; and (3) to develop a genetic screen in yeast to identify methotrexate-resistant variants of ATIC.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01CA075118-04
Application #
6173090
Study Section
Subcommittee G - Education (NCI)
Program Officer
Eckstein, David J
Project Start
1997-09-01
Project End
2002-08-31
Budget Start
2000-09-01
Budget End
2001-08-31
Support Year
4
Fiscal Year
2000
Total Cost
$145,019
Indirect Cost

  Institution

Name
Pennsylvania State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
003403953
City
University Park
State
PA
Country
United States
Zip Code
16802

  Publications

Arnold, Jamie J; Gohara, David W; Cameron, Craig E (2004) Poliovirus RNA-dependent RNA polymerase (3Dpol): pre-steady-state kinetic analysis of ribonucleotide incorporation in the presence of Mn2+. Biochemistry 43:5138-48
Arnold, Jamie J; Cameron, Craig E (2004) Poliovirus RNA-dependent RNA polymerase (3Dpol): pre-steady-state kinetic analysis of ribonucleotide incorporation in the presence of Mg2+. Biochemistry 43:5126-37
Gohara, David W; Arnold, Jamie J; Cameron, Craig E (2004) Poliovirus RNA-dependent RNA polymerase (3Dpol): kinetic, thermodynamic, and structural analysis of ribonucleotide selection. Biochemistry 43:5149-58
Pathak, Harsh B; Ghosh, Saikat Kumar B; Roberts, Allan W et al. (2002) Structure-function relationships of the RNA-dependent RNA polymerase from poliovirus (3Dpol). A surface of the primary oligomerization domain functions in capsid precursor processing and VPg uridylylation. J Biol Chem 277:31551-62
Crotty, Shane; Cameron, Craig; Andino, Raul (2002) Ribavirin's antiviral mechanism of action: lethal mutagenesis? J Mol Med 80:86-95
Maag, D; Castro, C; Hong, Z et al. (2001) Hepatitis C virus RNA-dependent RNA polymerase (NS5B) as a mediator of the antiviral activity of ribavirin. J Biol Chem 276:46094-8
Wei, L; Huhn, J S; Mory, A et al. (2001) Proteinase-polymerase precursor as the active form of feline calicivirus RNA-dependent RNA polymerase. J Virol 75:1211-9
Hong, Z; Cameron, C E; Walker, M P et al. (2001) A novel mechanism to ensure terminal initiation by hepatitis C virus NS5B polymerase. Virology 285:6-11
Tackett, A J; Wei, L; Cameron, C E et al. (2001) Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex. Nucleic Acids Res 29:565-72
Crotty, S; Maag, D; Arnold, J J et al. (2000) The broad-spectrum antiviral ribonucleoside ribavirin is an RNA virus mutagen. Nat Med 6:1375-9

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