Abstract

This proposal focused on the structural and functional characterization of enzymes involved in the biological synthesis of unusual natural products. A DNA-based approach to natural products, an approach where biosynthetic gene clusters are captured and heterologously expressed in host organisms has proven to be a powerful method for discovering new biologically active small molecules, new biosynthetic enzymes with unusual activity, and new signaling paradigms for small molecule-based signaling. The work proposed follows up on each of these themes and also proposes colony hybridization-based approaches to identifying potentially useful DNA sequences prior to heterologous expression. A genome wide scan with a synthetic library that amplifies the molecular diversity of the heterologously expressed libraries is also proposed. This scan, if successful, would provide functional annotation for the endogenous function of previously characterized small molecules. Finally, some polypeptides with antibiotic activity will be investigated.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA059021-13
Application #
6855094
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Lees, Robert G
Project Start
1992-07-10
Project End
2007-12-31
Budget Start
2005-02-01
Budget End
2006-01-31
Support Year
13
Fiscal Year
2005
Total Cost
$305,100
Indirect Cost

  Institution

Name
Harvard University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Ramadhar, Timothy R; Beemelmanns, Christine; Currie, Cameron R et al. (2014) Bacterial symbionts in agricultural systems provide a strategic source for antibiotic discovery. J Antibiot (Tokyo) 67:53-8
Wieland Brown, Laura C; Acker, Michael G; Clardy, Jon et al. (2009) Thirteen posttranslational modifications convert a 14-residue peptide into the antibiotic thiocillin. Proc Natl Acad Sci U S A 106:2549-53
Fischbach, Michael A; Walsh, Christopher T; Clardy, Jon (2008) The evolution of gene collectives: How natural selection drives chemical innovation. Proc Natl Acad Sci U S A 105:4601-8
Mazitschek, Ralph; Patel, Vishal; Wirth, Dyann F et al. (2008) Development of a fluorescence polarization based assay for histone deacetylase ligand discovery. Bioorg Med Chem Lett 18:2809-12
Junker, Lauren M; Clardy, Jon (2007) High-throughput screens for small-molecule inhibitors of Pseudomonas aeruginosa biofilm development. Antimicrob Agents Chemother 51:3582-90
Schmitz, Katja; Haggarty, Stephen J; McPherson, Olivia M et al. (2007) Detecting binding interactions using microarrays of natural product extracts. J Am Chem Soc 129:11346-7
Lautenschlager, Catherine; Leal, Walter S; Clardy, Jon (2007) Bombyx mori pheromone-binding protein binding nonpheromone ligands: implications for pheromone recognition. Structure 15:1148-54
Fischbach, Michael A; Clardy, Jon (2007) One pathway, many products. Nat Chem Biol 3:353-5
Clardy, Jon; Brady, Sean F (2007) Cyclic AMP directly activates NasP, an N-acyl amino acid antibiotic biosynthetic enzyme cloned from an uncultured beta-proteobacterium. J Bacteriol 189:6487-9
Schroeder, Frank C; Gibson, Donna M; Churchill, Alice C L et al. (2007) Differential analysis of 2D NMR spectra: new natural products from a pilot-scale fungal extract library. Angew Chem Int Ed Engl 46:901-4

Showing the most recent 10 out of 28 publications