Transcription from the chiC promoter, directing expression of the chitinase gene, chiC, in Streptomyces coelicolor, was analyzed using xylE reporter gene and high-resolution S1-nuclease mapping. The transcription from the chiC promoter was induced by chitin, and this induction was dramatically reduced in the S. coelicolor chiR-disrupted strain. This indicated a dependence of chiC expression upon the chiR gene encoding a response regulator protein. To investigate this relationship, the S. coelicolor ChiR was overproduced using Escherichia coli T7 RNA polymerase expression system. However, gel mobility shift-assay with such a purified ChiR showed no binding in the chiC promoter region, which indicates a lack of specific phosphorylation of E. coli overproduced ChiR that is necessary for DNA-binding activity of response regulators.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• chitinasy genetika   metabolismus   MeSH
• endonukleasy specifické pro jednořetězcové nukleové kyseliny MeSH
• Escherichia coli enzymologie   genetika   MeSH
• genetická transkripce * MeSH
• molekulární sekvence - údaje MeSH
• promotorové oblasti (genetika) MeSH
• regulace genové exprese u bakterií * MeSH
• restrikční mapování MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• Streptomyces enzymologie   genetika   MeSH
• transkripční faktory * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální proteiny MeSH
• ChiR protein, Streptomyces MeSH Prohlížeč
• chitinase C-1 MeSH Prohlížeč
• chitinasy MeSH
• endonukleasy specifické pro jednořetězcové nukleové kyseliny MeSH
• transkripční faktory * MeSH

Using the method for the identification of promoters recognized by the sporulation specific sigma factor (sigma F), we identified a positive 950 bp Sau3AI DNA fragment in Streptomyces coelicolor A3(2). High-resolution S1-nuclease mapping identified a potential promoter, PF35, in the E. coli two-plasmid system similar to the consensus sequence of Bacillus subtilis promoters recognized by the general stress-response sigma factor (sigma B). However, the putative sigF-dependent promoter, PF35, was inactive in S. coelicolor in the course of differentiation, and it was located divergently in the promoter region directing expression of the chiC gene encoding chitinase. Sequence analysis of the region potentially governed by PF35 revealed two translationally coupled genes encoding proteins similar to bacterial two-component regulatory systems, and with the highest similarity to the two-component system chiS, chiR, regulating chitinase activity in Streptomyces thermoviolaceus. However, the genes had a divergent orientation with respect to the PF35 promoter. Disruption of the S. coelicolor chiR gene appeared to have no obvious effect on growth, morphology, differentiation, and production of pigmented antibiotic actinorhodin and undecylprodigiosin. Moreover, the chiR disruption did not affect the overall chitinase activity.

• MeSH
• bakteriální proteiny chemie   genetika   metabolismus   MeSH
• chitinasy genetika   metabolismus   MeSH
• delece genu MeSH
• klonování DNA * MeSH
• molekulární sekvence - údaje MeSH
• operon MeSH
• promotorové oblasti (genetika) MeSH
• proteinkinasy * MeSH
• regulace genové exprese u bakterií genetika   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• sigma faktor MeSH
• Streptomyces enzymologie   genetika   růst a vývoj   MeSH
• transkripční faktory * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální proteiny MeSH
• ChiR protein, Streptomyces MeSH Prohlížeč
• ChiS protein, Streptomyces thermoviolaceus MeSH Prohlížeč
• chitinase C-1 MeSH Prohlížeč
• chitinasy MeSH
• FliA protein, Bacteria MeSH Prohlížeč
• proteinkinasy * MeSH
• sigma faktor MeSH
• transkripční faktory * MeSH