title: Structural and functional analysis of the TetR-family transcriptional regulator SCO0332 from Streptomyces coelicolor.
abstract: SCO0332 protein is a putative TetR-family transcriptional regulator from Streptomyces coelicolor A3(2). The crystal structure of SCO0332 was determined at 2.25 A resolution by single-wavelength anomalous diffraction (SAD) phasing using the S atoms of the native protein. SCO0332 contains a helix-turn-helix (HTH) DNA-binding motif in its N-terminal region and forms a homodimer. The overall structure of SCO0332 shows significant similarity to other TetR-family regulators. A systematic evolution of ligands by exponential enrichment (SELEX) analysis indicated that SCO0332 has sequence-specific DNA-binding ability and determined the position of the operator element of SCO0332 on the chromosomal DNA of S. coelicolor. An electrophoretic mobility-shift assay (EMSA) showed that SCO0332 binds to the operator sequence upstream of the sco0330 gene, which encodes a putative short-chain oxidoreductase. These results suggest that SCO0332 is a transcriptional repressor that regulates sco0330 gene expression.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Ui
authors.initials: U
authors.lastname: Okada
authors.forename: Kazuya
authors.initials: K
authors.lastname: Kondo
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hayashi
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Watanabe
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Tamura
authors.forename: Isao
authors.initials: I
authors.lastname: Tanaka
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray / Q000379:methods; D024202:Electrophoretic Mobility Shift Assay; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D048372:Streptomyces coelicolor / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins