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Lincomycin is one of the most important antibiotics in clinical practice. To further understand the regulatory mechanism on lincomycin biosynthesis, we investigated a pleiotropic transcriptional regulator AdpA lin  in the lincomycin producer  Streptomyces lincolnensis  NRRL 2936. Deletion of  adpA   lin   (which generated Δ adpA   lin  ) interrupted lincomycin biosynthesis and impaired the morphological differentiation. We also found that putative AdpA binding sites were unusually scattered in the promoters of all the 8 putative operons in the lincomycin biosynthetic gene cluster (BGC). In Δ adpA   lin  , transcript levels of structural genes in 8 putative operons were decreased with varying degrees, and electrophoretic mobility shift assays (EMSAs) confirmed that AdpA lin  activated the overall putative operons via directly binding to their promoter regions. Thus, we speculated that the entire lincomycin biosynthesis is under the control of AdpA lin . Besides, AdpA lin  participated in lincomycin biosynthesis by binding to the promoter of  lmbU  which encoded a cluster sited regulator (CSR) LmbU of lincomycin biosynthesis. Results of qRT-PCR and catechol dioxygenase activity assay showed that AdpA lin  activated the transcription of  lmbU . In addition, AdpA lin  activated the transcription of the  bldA  by binding to its promoter, suggesting that AdpA lin  indirectly participated in lincomycin biosynthesis and morphological differentiation. Uncommon but understandable, AdpA lin  auto-activated its own transcription via binding to its own promoter region. In conclusion, we provided a molecular mechanism around the effect of AdpA lin  on lincomycin biosynthesis in  S. lincolnensis , and revealed a cascade regulation of lincomycin biosynthesis by AdpA lin , LmbU, and BldA.

AdpAlin, a Pleiotropic Transcriptional Regulator, Is Involved in the Cascade Regulation of Lincomycin Biosynthesis in Streptomyces lincolnensis