caa a22 4500 605502781 CHVBK 20210128100607.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6638-6 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6638-6 Identification of two novel regulatory genes involved in pristinamycin biosynthesis and elucidation of the mechanism for AtrA-p-mediated regulation in Streptomyces pristinaespiralis [Elektronische Daten] [Wenfang Wang, Jinzhong Tian, Lei Li, Mei Ge, Hong Zhu, Guosong Zheng, He Huang, Lijun Ruan, Weihong Jiang, Yinhua Lu] In this study, using a transposon-based strategy, two novel regulatory genes were identified as being involved in the biosynthesis of both pristinamycin I (PI) and II (PII) in Streptomyces pristinaespiralis, including a TetR-family regulatory gene atrA-p (SSDG_00466) and an orphan histidine kinase gene SSDG_02492. The mechanism by which AtrA-p exerted a positive role in pristinamycin production was elucidated. We showed that deletion of atrA-p resulted in a delayed production of both PI and PII as well as reduced PII production. Transcriptional analysis integrated with electrophoretic mobility shift assays (EMSAs) demonstrated that AtrA-p played a positive role in pristinamycin production by directly activating the transcription of two cluster-situated regulatory genes, spbR and papR5, which encode a γ-butyrolactone receptor protein and a TetR-family repressor, respectively. The precise AtrA-p-binding sites upstream of these two targets were determined, which allowed the identification of a relatively conserved binding motif comprising two 5-nt inverted repeats separated by a variable 5-nt sequence (5′-GGAAT-n5-ATTCC-3′) possibly required for the regulation of AtrA-like regulators in Streptomyces. Base substitutions of the AtrA-p-binding sites on the genome caused similar decreases in spbR and papR5 transcription as those observed in ∆atrA-p. Taken together, herein, a novel mechanism for AtrA-dependent regulation of antibiotic biosynthesis was revealed in S. pristinaespiralis, which is distinct from those of its homologs, AtrA-c from Streptomyces coelicolor, AtrA-g from Streptomyces griseus, and AtrA from Streptomyces roseosporus that perform their effects in antibiotic biosynthesis directly via pathway-specific activator genes or the biosynthetic structural genes. Springer-Verlag Berlin Heidelberg, 2015 Streptomyces pristinaespiralis nationallicence Pristinamycin nationallicence Regulation nationallicence Transposon nationallicence AtrA nationallicence Wang Wenfang Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Tian Jinzhong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Li Lei Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Ge Mei Shanghai Laiyi Center for Biopharmaceuticals R&D, 201203, Shanghai, People's Republic of China aut Zhu Hong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Zheng Guosong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Huang He Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Ruan Lijun Shanghai Laiyi Center for Biopharmaceuticals R&D, 201203, Shanghai, People's Republic of China aut Jiang Weihong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Lu Yinhua Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7151-7164 0175-7598 99:17<7151 2015 99 253 https://doi.org/10.1007/s00253-015-6638-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6638-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Wenfang Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Tian Jinzhong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Li Lei Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Ge Mei Shanghai Laiyi Center for Biopharmaceuticals R&D, 201203, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Zhu Hong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Guosong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Huang He Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Ruan Lijun Shanghai Laiyi Center for Biopharmaceuticals R&D, 201203, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Jiang Weihong Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Lu Yinhua Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7151-7164 0175-7598 99:17<7151 2015 99 253