TY - JOUR
T1 - The biosynthetic gene cluster for the antitumor drug bleomycin from Streptomyces verticillus ATCC15003 supporting functional interactions between nonribosomal peptide synthetases and a polyketide synthase
AU - Du, Liangcheng
AU - Sánchez, César
AU - Chen, Mei
AU - Edwards, Daniel J.
AU - Shen, Ben
N1 - Funding Information:
We thank Dr. M. Calcutt and Prof. F. Schmidt, University of Missouri–Columbia, for clones of the blmAB locus, Dr. T. O’Hare, Oregon State University, for advice on constructing genomic library in pOJ446, Dr. J. Wu at the Cancer Center, University of California, Davis, for EIS–MS analysis, and Prof. C.R. Hutchinson, University of Wisconsin, Madison, for helpful discussion and constant encouragement. This work was supported in part by an Institutional Research Grant from the American Cancer Society and the School of Medicine, University of California, Davis; the National Institutes of Health Grant AI40475; and the Searle Scholars Program/The Chicago Community Trust. C.S. was supported in part by a postdoctoral fellowship from the Spanish Ministry of Education and Culture.
PY - 2000
Y1 - 2000
N2 - Background: The structural and catalytic similarities between modular nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) inspired us to search for a hybrid NRPS-PKS system. The antitumor drug bleomycin (BLM) is a natural hybrid peptide-polyketide metabolite, the biosynthesis of which provides an excellent opportunity to investigate intermodular communication between NRPS and PKS modules. Here, we report the cloning, sequencing, and characterization of the BLM biosynthetic gene cluster from Streptomyces verticillus ATCC15003. Results: A set of 30 genes clustered with the previously characterized blmAB resistance genes were defined by sequencing a 85-kb contiguous region of DNA from S. verticillus ATCC15003. The sequenced gene cluster consists of 10 NRPS genes encoding nine NRPS modules, a PKS gene encoding one PKS module, five sugar biosynthesis genes, as well as genes encoding other biosynthesis, resistance, and regulatory proteins. The substrate specificities of individual NRPS and PKS modules were predicted based on sequence analysis, and the amino acid specificities of two NRPS modules were confirmed biochemically in vitro. The involvement of the cloned genes in BLM biosynthesis was demonstrated by bioconversion of the BLM aglycones into BLMs in Streptomyces lividans expressing a part of the gene cluster. Conclusion: The blm gene cluster is characterized by a hybrid NRPS-PKS system, supporting the wisdom of combining individual NRPS and PKS modules for combinatorial biosynthesis. The availability of the blm gene cluster has set the stage for engineering novel BLM analogs by genetic manipulation of genes governing BLM biosynthesis and for investigating the molecular basis for intermodular communication between NRPS and PKS in the biosynthesis of hybrid peptide-polyketide metabolites.
AB - Background: The structural and catalytic similarities between modular nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) inspired us to search for a hybrid NRPS-PKS system. The antitumor drug bleomycin (BLM) is a natural hybrid peptide-polyketide metabolite, the biosynthesis of which provides an excellent opportunity to investigate intermodular communication between NRPS and PKS modules. Here, we report the cloning, sequencing, and characterization of the BLM biosynthetic gene cluster from Streptomyces verticillus ATCC15003. Results: A set of 30 genes clustered with the previously characterized blmAB resistance genes were defined by sequencing a 85-kb contiguous region of DNA from S. verticillus ATCC15003. The sequenced gene cluster consists of 10 NRPS genes encoding nine NRPS modules, a PKS gene encoding one PKS module, five sugar biosynthesis genes, as well as genes encoding other biosynthesis, resistance, and regulatory proteins. The substrate specificities of individual NRPS and PKS modules were predicted based on sequence analysis, and the amino acid specificities of two NRPS modules were confirmed biochemically in vitro. The involvement of the cloned genes in BLM biosynthesis was demonstrated by bioconversion of the BLM aglycones into BLMs in Streptomyces lividans expressing a part of the gene cluster. Conclusion: The blm gene cluster is characterized by a hybrid NRPS-PKS system, supporting the wisdom of combining individual NRPS and PKS modules for combinatorial biosynthesis. The availability of the blm gene cluster has set the stage for engineering novel BLM analogs by genetic manipulation of genes governing BLM biosynthesis and for investigating the molecular basis for intermodular communication between NRPS and PKS in the biosynthesis of hybrid peptide-polyketide metabolites.
KW - Biosynthesis
KW - Bleomycin
KW - Nonribosomal peptide synthetase
KW - Polyketide synthase
KW - Streptomyces verticillus
UR - http://www.scopus.com/inward/record.url?scp=0033835373&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033835373&partnerID=8YFLogxK
U2 - 10.1016/S1074-5521(00)00011-9
DO - 10.1016/S1074-5521(00)00011-9
M3 - Article
C2 - 11048953
AN - SCOPUS:0033835373
VL - 7
SP - 623
EP - 642
JO - Cell Chemical Biology
JF - Cell Chemical Biology
SN - 2451-9448
IS - 8
ER -