The goal of the proposed research is to study the functionality of the domains of the iterative type I enediyne C-1027 polyketide synthase (PKS). Conserved domains will be analysized by site-directed mutagenesis of the proposed active sites, and the mutants characterized by in vivo analysis. It is hypothesized the PKS contains an unusual acyl carrier protein (ACP) domain that requires a unique phosphopantetheinyl transferase (PPTase), which is proposed to also be a domain found within the PKS. The function of the putative ACP and PPTase domain will be analyzed by a thorough in vivo and in vitro analysis, which includes mutant generation of the proposed active sites. Finally, it is proposed that the regiochemistry of enediyne formation is controlled by the PKS. This will be tested by the creation of hybrid PKSs from C- 1027, which contains a 9-carbon enediyne core, and calicheamicin, which contains a 10-carbon enediyne core. The results will provide insights into the programming rules of enediyne biosynthesisand will allow for the generation of """"""""unnatural"""""""" natural enediynes with predicted chemistry.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32CA105984-01
Application #
6741023
Study Section
Special Emphasis Panel (ZRG1-F04 (20))
Program Officer
Lohrey, Nancy
Project Start
2004-05-01
Project End
2007-04-30
Budget Start
2004-05-01
Budget End
2005-04-30
Support Year
1
Fiscal Year
2004
Total Cost
$41,168
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Liu, Wen; Nonaka, Koichi; Nie, Liping et al. (2005) The neocarzinostatin biosynthetic gene cluster from Streptomyces carzinostaticus ATCC 15944 involving two iterative type I polyketide synthases. Chem Biol 12:293-302
Galm, Ute; Hager, Martin H; Van Lanen, Steven G et al. (2005) Antitumor antibiotics: bleomycin, enediynes, and mitomycin. Chem Rev 105:739-58