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However, the underlying mechanism of how it engages bacterial RNA polymerase and promoter DNA to finely respond to environmental stimuli is still elusive. Here, we present two cryo-EM structures of Rob-dependent transcription activation complex (Rob-TAC) comprising of Escherichia coli RNA polymerase (RNAP), Rob-regulated promoter and Rob in alternative conformations. The structures show that a single Rob engages RNAP by interacting with RNAP \u03b1CTD and \u03c370R4, revealing their generally important regulatory roles. Notably, by occluding \u03c370R4 from binding to -35 element, Rob specifically binds to the conserved Rob binding box through its consensus HTH motifs, and retains DNA bending by aid of the accessory acidic loop. More strikingly, our ligand docking and biochemical analysis demonstrate that the large Rob C-terminal domain (Rob CTD) shares great structural similarity with the global Gyrl-like domains in effector binding and allosteric regulation, and coordinately promotes formation of competent Rob-TAC. Altogether, our structural and biochemical data highlight the detailed molecular mechanism of Rob-dependent transcription activation, and provide favorable evidences for understanding the physiological roles of the other AraC\/XylS-family transcription factors.<\/jats:p>","DOI":"10.1093\/nar\/gkac433","type":"journal-article","created":{"date-parts":[[2022,5,9]],"date-time":"2022-05-09T07:10:00Z","timestamp":1652080200000},"page":"5974-5987","source":"Crossref","is-referenced-by-count":28,"title":["Structural basis of transcription activation by Rob, a pleiotropic AraC\/XylS family regulator"],"prefix":"10.1093","volume":"50","author":[{"given":"Jing","family":"Shi","sequence":"first","affiliation":[{"name":"Department of Pathogen Biology, School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine , Nanjing 210023 , China"},{"name":"Department of Pathology of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine , Hangzhou 310058 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fulin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathogen Biology, School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine , Nanjing 210023 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangfang","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pathogen Biology, School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine , Nanjing 210023 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathogen Biology, School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine , Nanjing 210023 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Xiong","sequence":"additional","affiliation":[{"name":"Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences , Beijing 100190 , China"},{"name":"School of Physics, University of Chinese Academy of Sciences , Beijing 100049 , China"},{"name":"Songshan Lake Materials Laboratory , Dongguan 523808 , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aijia","family":"Wen","sequence":"additional","affiliation":[{"name":"Department of Biophysics, Zhejiang University School of Medicine , Hangzhou 310058 , China"},{"name":"Department of Pathology of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine , Hangzhou 310058 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanling","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Pathogen Biology, School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine , Nanjing 210023 , 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