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Titolo:
Characterization and analysis of the PikD regulatory factor in the pikromycin biosynthetic pathway of Streptomyces venezuelae
Autore:
Wilson, DJ; Xue, YQ; Reynolds, KA; Sherman, DH;
Indirizzi:
Univ Minnesota, Dept Microbiol, Minneapolis, MN 55455 USA Univ Minnesota Minneapolis MN USA 55455 robiol, Minneapolis, MN 55455 USA Univ Minnesota, Biol Proc Technol Inst, Minneapolis, MN 55455 USA Univ Minnesota Minneapolis MN USA 55455 l Inst, Minneapolis, MN 55455 USA Virginia Commonwealth Univ, Dept Med Chem, Richmond, VA 23219 USA VirginiaCommonwealth Univ Richmond VA USA 23219 , Richmond, VA 23219 USA
Titolo Testata:
JOURNAL OF BACTERIOLOGY
fascicolo: 11, volume: 183, anno: 2001,
pagine: 3468 - 3475
SICI:
0021-9193(200106)183:11<3468:CAAOTP>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
DNA-BINDING PROTEIN; GENE-CLUSTER; COELICOLOR A3(2); SECONDARY METABOLISM; POLYKETIDE SYNTHASE; NUCLEOTIDE-SEQUENCE; ESCHERICHIA-COLI; CLONING VECTORS; RAPAMYCIN; FAMILY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Wilson, DJ Univ Minnesota, Dept Microbiol, Mayo Mail Code 196,420 DelawareSt SE, Minneapolis, MN 55455 USA Univ Minnesota Mayo Mail Code 196,420 Delaware St SE Minneapolis MN USA 55455
Citazione:
D.J. Wilson et al., "Characterization and analysis of the PikD regulatory factor in the pikromycin biosynthetic pathway of Streptomyces venezuelae", J BACT, 183(11), 2001, pp. 3468-3475

Abstract

The Streptomyces venezuelae pikD gene from the pikromycin biosynthetic cluster was analyzed, and its deduced product (PikD) was found to have amino acid sequence homology with a small family of bacterial regulatory proteins. Database comparisons revealed two hypothetical domains, including an N-terminal triphosphate-binding domain and a C-terminal helix-turn-helix DNA-binding motif, Analysis of PikD was initiated by deletion of the correspondinggene (pikD) from the chromosome of S. venezuelae, resulting in complete loss of antibiotic production. Complementation by a plasmid carrying pikD restored macrolide biosynthesis, demonstrating that PikD is a positive regulator. Mutations were made in the predicted nucleotide triphosphate-binding domain, confirming the active-site amino acid residues of the Walker A and B motifs. Feeding of macrolide intermediates was carried out to gauge the points of operon control by PikD. Although the pikD mutant strain was unable to convert macrolactones (10-deoxymethynolide and narbonolide) to glycosylated products, macrolide intermediates (YC-17 and narbomycin) were hydroxylated with high efficiency. To study further the control of biosynthesis, presumed promoter regions from pik cluster loci were linked to the xylE reporter and placed in S. venezuelae wild-type and pikD mutant strains. This analysis demonstrated that PikD-mediated transcriptional regulation occurs at promoters controlling expression of pikRII, pikAI, and desI but not those controlling pikRI or pikC.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 02:14:06