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. 2021 Feb 9;64(11):1949–1963. doi: 10.1007/s11427-020-1892-3

Molecular mechanism of mureidomycin biosynthesis activated by introduction of an exogenous regulatory gene ssaA into Streptomyces roseosporus

Ning Liu 1,2, Hanye Guan 1,2, Guoqing Niu 3, Lingjuan Jiang 1,4, Yue Li 1, Jihui Zhang 1, Jine Li 1, Huarong Tan 1,2,✉
PMCID: PMC7880210  PMID: 33580428

Abstract

Mureidomycins (MRDs), a group of unique uridyl-peptide antibiotics, exhibit antibacterial activity against the highly refractory pathogen Pseudomonas aeruginosa. Our previous study showed that the cryptic MRD biosynthetic gene cluster (BGC) mrd in Streptomyces roseosporus NRRL 15998 could not be activated by its endogenous regulator 02995 but activated by an exogenous activator SsaA from sansanmycin’s BGC ssa of Streptomyces sp. strain SS. Here we report the molecular mechanism for this inexplicable regulation. EMSAs and footprinting experiments revealed that SsaA could directly bind to a 14-nt palindrome sequence of 5′-CTGRCNNNNGTCAG-3′ within six promoter regions of mrd. Disruption of three representative target genes (SSGG-02981, SSGG-02987 and SSGG-02994) showed that the target genes directly controlled by SsaA were essential for MRD production. The regulatory function was further investigated by replacing six regions of SSGG-02995 with those of ssaA. Surprisingly, only the replacement of 343–450 nt fragment encoding the 115–150 amino acids (AA) of SsaA could activate MRD biosynthesis. Further bioinformatics analysis showed that the 115–150 AA situated between two conserved domains of SsaA. Our findings significantly demonstrate that constitutive expression of a homologous exogenous regulatory gene is an effective strategy to awaken cryptic biosynthetic pathways in Streptomyces.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s11427-020-1892-3 and is accessible for authorized users.

Keywords: mureidomycin, transcriptional activation, exogenous regulator—SsaA, cryptic gene cluster, Streptomyces roseosporus, Pseudomonas aeruginosa, antibiotics

Supplementary Materials

11427_2020_1892_MOESM1_ESM.pdf (1.1MB, pdf)

Supplementary material, approximately 1.13 MB.

Acknowledgements

This work was supported by the National Key Research and Development Program of China (2020YFA0907800 and 2018YFA0901900) and the National Natural Science Foundation of China (81773615, 31771378 and 31800029). We are very grateful to Drs. Wang Wenxi, Liu Xiang and Zheng Jiazhen (Institute of Microbiology, Chinese Academy of Sciences, Beijing, China) for their technical advice in EMSA and DNase I footprinting experiments.

Compliance and ethics The author(s) declare that they have no conflict of interest.

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Supplementary Materials

11427_2020_1892_MOESM1_ESM.pdf (1.1MB, pdf)

Supplementary material, approximately 1.13 MB.

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