Biosynthetic pathway and gene cluster analysis of curacin A, an antitubulin natural product from the tropical marine cyanobacterium Lyngbya majuscula.
Curacin A (1) is a potent cancer cell toxin obtained from strains of the tropical marine cyanobacterium Lyngbya majuscula found in Curaçao. Its structure is unique in that it contains the sequential positioning of a thiazoline and cyclopropyl ring, and it exerts its potent cell toxicity through interaction with the colchicine drug binding site on microtubules. A series of stable isotope-labeled precursors were fed to cultures of curacin A-producing strains and, following NMR analysis, allowed determination of the metabolic origin of all atoms in the natural product (one cysteine, 10 acetate units, two S-adenosyl methionine-derived methyl groups) as well as several unique mechanistic insights. Moreover, these incorporation experiments facilitated an effective gene cloning strategy that allowed identification and sequencing of the approximately 64 kb putative curacin A gene cluster. The metabolic system is comprised of a nonribosomal peptide synthetase (NRPS) and multiple polyketide synthases (PKSs) and shows a very high level of collinearity between genes in the cluster and the predicted biochemical steps required for curacin biosynthesis. Unique features of the cluster include (1) all but one of the PKSs are monomodular multifunctional proteins, (2) a unique gene cassette that contains an HMG-CoA synthase likely responsible for formation of the cyclopropyl ring, and (3) a terminating motif that is predicted to function in both product release and terminal dehydrative decarboxylation.
Chang
Zunxue
Z
Department of Medicinal Chemistry, College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109, USA.
Sitachitta
Namthip
N
Rossi
James V
JV
Roberts
Mary Ann
MA
Flatt
Patricia M
PM
Jia
Junyong
J
Sherman
David H
DH
Gerwick
William H
WH
D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000378:metabolism* / Q000494:pharmacology*; D000458:Cyanobacteria / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003521:Cyclopropanes / Q000378:metabolism* / Q000494:pharmacology*; D008387:Marine Toxins / Q000378:metabolism* / Q000494:pharmacology*; D008938:Mitosis / Q000187:drug effects*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D016415:Sequence Alignment; D013844:Thiazoles / Q000378:metabolism* / Q000494:pharmacology*; D050257:Tubulin Modulators
D000970:Antineoplastic Agents; D003521:Cyclopropanes; D008387:Marine Toxins; D013844:Thiazoles; D050257:Tubulin Modulators; C095843:curacin A