An ABC transporter involved in the control of streptomycin production in Streptomyces griseus

• Published in: FEMS microbiology letters 2016-07, Vol.363 (14), p.fnw149
• Main Authors: Takano, Hideaki, Toriumi, Naoe, Hirata, Mariko, Amano, Taisuke, Ohya, Takaaki, Shimada, Reona, Kusada, Hiroyuki, Amano, Sho-ichi, Matsuda, Ko-ichi, Beppu, Teruhiko, Ueda, Kenji
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biosynthesis; Cloning, Molecular; Drug therapy; Gene Order; Genes; Genetic Loci; Microbiology; Mutation; Plasmids - genetics; Protein Interaction Domains and Motifs; Streptomyces griseus - genetics; Streptomyces griseus - metabolism; Streptomycin - biosynthesis
• ISSN: 1574-6968; 0378-1097; 1574-6968
• DOI: 10.1093/femsle/fnw149

We screened for a gene that inhibits streptomycin production in Streptomyces griseus when it is introduced on a high-copy-number plasmid pIJ702, and obtained a plasmid pKM545. The introduction of pKM545 abolished streptomycin production on all media tested including YMP-sugar and Nutrient broth. S1 protection analysis demonstrated that the introduction of this plasmid downregulated the transcriptional activity of the promoter preceding strR, the pathway-specific transcriptional regulator for streptomycin biosynthesis. The 2.8-kb BamHI fragment cloned onto pKM545 contained two coding sequences SGR_5442 and 5443. These coding sequences and the two downstream ones (SGR_5444 and 5445) constituted a possible operon structure designated to be rspABCD (regulation of streptomycin production). RspB and RspC exhibited a marked similarity with an ATP-binding domain and a membrane-associating domain of an ABC-2 type transporter, respectively, suggesting that the Rsp proteins comprise a membrane exporter. The gene cluster consisting of the rsp operon and the upstream divergent small coding sequence (SGR_5441) was widely distributed to Streptomyces genome. An rspB mutant of S. griseus produced 3-fold streptomycin of the parental strain in YMP liquid medium. The evidence implies that the Rsp translocator is involved in the export of a substance that specifies the expression level of streptomycin biosynthesis genes in S. griseus. An ABC transporter involved in the control of streptomycin production in Streptomyces griseus.