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Journal of Bacteriology, March 2004, p. 1345-1354, Vol. 186, No. 5
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.5.1345-1354.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

In Vivo Analysis of the Regulatory Genes in the Nystatin Biosynthetic Gene Cluster of Streptomyces noursei ATCC 11455 Reveals Their Differential Control Over Antibiotic Biosynthesis

Olga N. Sekurova,1 Trygve Brautaset,1 Håvard Sletta,2 Sven E. F. Borgos,1 Øyvind M. Jakobsen,1 Trond E. Ellingsen,2 Arne R. Strøm,1 Svein Valla,1 and Sergey B. Zotchev1*

Department of Biotechnology, Norwegian University of Science and Technology, N-7491 Trondheim,1 SINTEF Industrial Biotechnology, SINTEF, N-7034 Trondheim, Norway2

Received 10 September 2003/ Accepted 21 November 2003

Six putative regulatory genes are located at the flank of the nystatin biosynthetic gene cluster in Streptomyces noursei ATCC 11455. Gene inactivation and complementation experiments revealed that nysRI, nysRII, nysRIII, and nysRIV are necessary for efficient nystatin production, whereas no significant roles could be demonstrated for the other two regulatory genes. To determine the in vivo targets for the NysR regulators, chromosomal integration vectors with the xylE reporter gene under the control of seven putative promoter regions upstream of the nystatin structural and regulatory genes were constructed. Expression analyses of the resulting vectors in the S. noursei wild-type strain and regulatory mutants revealed that the four regulators differentially affect certain promoters. According to these analyses, genes responsible for initiation of nystatin biosynthesis and antibiotic transport were the major targets for regulation. Data from cross-complementation experiments showed that nysR genes could in some cases substitute for each other, suggesting a functional hierarchy of the regulators and implying a cascade-like mechanism of regulation of nystatin biosynthesis.

* Corresponding author. Mailing address: Department of Biotechnology, Norwegian University of Science and Technology, N-7491 Trondheim, Norway. Phone: 47 73 59 86 79. Fax: 47 73 59 12 83. E-mail: sergey.zotchev{at}biotech.ntnu.no.

Journal of Bacteriology, March 2004, p. 1345-1354, Vol. 186, No. 5
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.5.1345-1354.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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