Direct binding of TFE&#945; opens DNA binding cleft of RNA polymerase.

Jun, Sung-Hoon; Hyun, Jaekyung; Cha, Jeong Seok; Kim, Hoyoung; Bartlett, Michael S; Cho, Hyun-Soo; Murakami, Katsuhiko S

Direct binding of TFEα opens DNA binding cleft of RNA polymerase.

Jun, Sung-Hoon; Hyun, Jaekyung; Cha, Jeong Seok; Kim, Hoyoung; Bartlett, Michael S; Cho, Hyun-Soo; Murakami, Katsuhiko S.

Afiliación

Jun SH; Electron Microscopy Research Center, Korea Basic Science Institute, Chungcheongbukdo, 28119, Republic of Korea. jsh100@kbsi.re.kr.
Hyun J; Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea. jsh100@kbsi.re.kr.
Cha JS; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, 16802, USA. jsh100@kbsi.re.kr.
Kim H; Electron Microscopy Research Center, Korea Basic Science Institute, Chungcheongbukdo, 28119, Republic of Korea.
Bartlett MS; Molecular Cryo-electron Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, Okinawa, 904-0495, Japan.
Cho HS; Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea.
Murakami KS; Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea.

Nat Commun ; 11(1): 6123, 2020 11 30.

Article en En | MEDLINE | ID: mdl-33257704

RESUMEN

Opening of the DNA binding cleft of cellular RNA polymerase (RNAP) is necessary for transcription initiation but the underlying molecular mechanism is not known. Here, we report on the cryo-electron microscopy structures of the RNAP, RNAP-TFEα binary, and RNAP-TFEα-promoter DNA ternary complexes from archaea, Thermococcus kodakarensis (Tko). The structures reveal that TFEα bridges the RNAP clamp and stalk domains to open the DNA binding cleft. Positioning of promoter DNA into the cleft closes it while maintaining the TFEα interactions with the RNAP mobile modules. The structures and photo-crosslinking results also suggest that the conserved aromatic residue in the extended winged-helix domain of TFEα interacts with promoter DNA to stabilize the transcription bubble. This study provides a structural basis for the functions of TFEα and elucidates the mechanism by which the DNA binding cleft is opened during transcription initiation in the stalk-containing RNAPs, including archaeal and eukaryotic RNAPs.

Asunto(s)

Proteínas de Unión al ADN/química; Proteínas de Unión al ADN/metabolismo; ARN Polimerasas Dirigidas por ADN/química; ARN Polimerasas Dirigidas por ADN/metabolismo; ADN/metabolismo; Thermococcus/enzimología; Secuencia de Aminoácidos; Microscopía por Crioelectrón; Cristalografía por Rayos X; Proteínas de Unión al ADN/genética; ARN Polimerasas Dirigidas por ADN/genética; Modelos Moleculares; Politetrafluoroetileno; Regiones Promotoras Genéticas; Unión Proteica; Conformación Proteica; Análisis de Secuencia de Proteína; Thermococcus/genética; Transcripción Genética

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Polimerasas Dirigidas por ADN / ADN / Thermococcus / Proteínas de Unión al ADN Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2020 Tipo del documento: Article Pais de publicación: Reino Unido

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