title: kasT gene of Streptomyces kasugaensis M338-M1 encodes a DNA-binding protein which binds to intergenic region of kasU-kasJ in the kasugamycin biosynthesis gene cluster.
abstract: We previously reported that a 4.2 kb SacI-EcoRI DNA region from Streptomyces kasugaensis M338-M1, a kasugamycin (KSM) producer, included KSM transporter genes (kasKLM). As an extension of that study, a 3.7 kb Psti-SacI DNA region, located at 1.5 approximately 5.2 kb upstream of kasK, was cloned and sequenced, revealing three complete open reading frames, designated kasT, kasU and kasJ. The kasJ gene encodes a protein (KasJ) with a conserved dinucleotide (FAD)-binding motif Homology search for KasJ showed its similarity to NADH: N-amidino-scyllo-inosamine oxidoreductase (StsB) which is involved in biosynthesis of the streptidine moiety of streptomycin (SM) in S. griseus. The kasT gene encodes a DNA-binding protein (KasT), including a helix-turn-helix motif near the center of the sequence. This protein is similar in structure to a pathway-specific activator protein (StrR) that plays a role in regulating the SM biosynthesis gene cluster of S. griseus. A fusion protein (Trx-KasT) clearly showed DNA binding activity with the intergenic region of kasU-kasJ, suggesting that KasT is a pathway-specific regulator of the KSM biosynthesis gene cluster.
authors.affiliation: Showa Pharmaceutical University, 3-3165 Higashi Tamagawagakuen, Machida-shi, Tokyo 194-8543, Japan. ikeno@ac.shoyaku.ac.jp
authors.forename: Souichi
authors.initials: S
authors.lastname: Ikeno
authors.forename: Daisuke
authors.initials: D
authors.lastname: Aoki
authors.forename: Koji
authors.initials: K
authors.lastname: Sato
authors.forename: Masa
authors.initials: M
authors.lastname: Hamada
authors.forename: Makoto
authors.initials: M
authors.lastname: Hori
authors.forename: Kayoko S
authors.initials: KS
authors.lastname: Tsuchiya
mesh_terms: D000595:Amino Acid Sequence; D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D019077:Helix-Turn-Helix Motifs; D008969:Molecular Sequence Data; D005810:Multigene Family; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C098255:StrR protein, Streptomyces griseus; D014157:Transcription Factors; C472174:kasT protein, Streptomyces kasugaensis; D005182:Flavin-Adenine Dinucleotide; C100305:kasugamycin