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J. Biol. Chem., Vol. 279, Issue 5, 3885-3892, January 30, 2004

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RNA Polymerase Mutation Activates the Production of a Dormant Antibiotic 3,3'-Neotrehalosadiamine via an Autoinduction Mechanism in Bacillus subtilis^*

Takashi Inaoka, Kosaku Takahashi, Hiroshi Yada, Mitsuru Yoshida, and Kozo Ochi¶

From the Microbial Function Laboratory and Molecular Elucidation Laboratory, National Food Research Institute, Tsukuba, Ibaraki 305-8642, Japan

Bacillus and Streptomyces species possess the ability to produce a variety of commercially important metabolites and extracellular enzymes. We previously demonstrated that antibiotic production in Streptomyces coeli-color A3(2) and Streptomyces lividans can be enhanced by RNA polymerase (RNAP) mutations selected for the rifampicin-resistant (Rif^r) phenotype. Here, we have shown that the introduction of a certain Rif^r rpoB mutation into a B. subtilis strain resulted in cells that overproduce an aminosugar antibiotic 3,3'-neotrehalosadiamine (NTD), the production of which is dormant in the wild-type strain. Mutational and recombinant gene expression analyses have revealed a polycistronic gene ntdABC (formally yhjLKJ) and a monocistronic gene ntdR (formally yhjM) as the NTD biosynthesis operon and a positive regulator for ntdABC, respectively. Analysis of transcriptional fusions to a lacZ reporter revealed that NTD acts as an autoinducer for its own biosynthesis genes via NtdR protein. Our results also showed that the Rif^r rpoB mutation causes an increase in the activity of ^A-dependent promoters including ntdABC promoter. Therefore, we propose that unlike the wild-type RNAP, the mutant RNAP efficiently recognized the ^A-dependent promoters, resulting in the dramatic activation of the NTD biosynthesis pathway by an autoinduction mechanism.

Received for publication, September 8, 2003 , and in revised form, November 11, 2003.

^* This work was supported by a grant from the Organized Research Combination System of the Science and Technology Agency of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Tel.: 81-29-838-8125; Fax: 81-29-838-7996; E-mail:kochi{at}affrc.go.jp.

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