An Alternative σ Factor, σ⁸, Controls Avermectin Production and Multiple Stress Responses in Streptomyces avermitilis

Alternative σ factors in bacteria redirect RNA polymerase to recognize alternative promoters, thereby facilitating coordinated gene expression necessary for adaptive responses. The gene sig8 (sav_741) in Streptomyces avermitilis encodes an alternative σ factor, σ⁸, highly homologous to σ^B in Streptomyces coelicolor. Studies reported here demonstrate that σ⁸ is an important regulator of both avermectin production and stress responses in S. avermitilis. σ⁸ inhibited avermectin production by indirectly repressing expression of cluster-situated activator gene aveR, and by directly initiating transcription of its downstream gene sav_742, which encodes a direct repressor of ave structural genes. σ⁸ had no effect on cell growth or morphological differentiation under normal growth conditions. Growth of a sig8-deletion mutant was less than that of wild-type strain on YMS plates following treatment with heat, H₂O₂, diamide, NaCl, or KCl. sig8 transcription was strongly induced by these environmental stresses, indicating response by σ⁸ itself. A series of σ⁸-dependent genes responsive to heat, oxidative and osmotic stress were identified by EMSAs, qRT-PCR and in vitro transcription experiments. These findings indicate that σ⁸ plays an important role in mediating protective responses to various stress conditions by activating transcription of its target genes. Six σ⁸-binding promoter sequences were determined and consensus binding sequence BGVNVH-N₁₅-GSNNHH (B: C, T or G, V: A, C or G, S: C or G, H: A, C or T, N: any nucleotide) was identified, leading to prediction of the σ⁸ regulon. The list consists of 940 putative σ⁸ target genes, assignable to 17 functional groups, suggesting the wide range of cellular functions controlled by σ⁸ in S. avermitilis.