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

• Published in: Journal of antibiotics Vol. 55; no. 12; pp. 1053 - 1062
• Main Authors: IKENO, SOUICHI, AOKI, DAISUKE, SATO, KOJI, HAMADA, MASA, HORI, MAKOTO, TSUCHIYA, KAYOKO S.
• Format: Journal Article
• Language: English
• Published: Tokyo JAPAN ANTIBIOTICS RESEARCH ASSOCIATION 2002; Japan Antibiotics Research Association
• Subjects: Amino Acid Sequence; Aminoglycosides; Anti-Bacterial Agents - biosynthesis; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Base Sequence; Binding Sites; Biological and medical sciences; Biotechnology; Cloning, Molecular; DNA, Intergenic; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Flavin-Adenine Dinucleotide - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial; Genetic engineering; Genetic technics; Helix-Turn-Helix Motifs; Methods. Procedures. Technologies; Molecular cloning; Molecular Sequence Data; Multigene Family; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Streptomyces - genetics; Streptomyces - metabolism; Transcription Factors - chemistry; Transcription Initiation Site; Transcription, Genetic; Gene cluster; Streptomycetaceae; Nucleotide sequence; DNA binding protein; Streptomyces; Biosynthesis; Open reading frame; Actinomycetales; Antibiotic; Production; Bacteria; Actinomycetes; Molecular cloning; Fusion protein
• ISSN: 0021-8820; 1881-1469
• DOI: 10.7164/antibiotics.55.1053

We previously reported that a 4.2kb 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 3.7kb PstI-SacI DNA region, located at 1.5-5.2kb 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.