Analysis of the draft sequence of the genome of teicoplanin producer Actinoplanes teichomyceticus (NRRL-B16726) led to identification of several genes encoding AraC-family regulators that resemble AdpA, master regulator of transcription in Streptomyces. We elucidated possible regulatory functions of one of the identified genes, adpA19(at), most similar to archetypal adpA from model Streptomyces species, in a series of expression experiments. Introduction of adpA19 at under control of its own promoter on moderate copy number vector pKC1139 into NRRL-B16726 had no influence on antibiotic production and sporulation. Introduction of adpA19 at into Streptomyces coelicolor M145 and several S. ghanaensis strains had major influence on antibiotic production by these bacteria. Finally, adpA19 at expression in a set of soil actinomycete isolates led to induction of synthesis of antibiotic compounds. Our data point to pleiotropic regulatory role of adpA19(at), warranting its use as a tool to manipulate secondary metabolome of actinomycetes.
Low molecular weight signaling compounds (LMWC) are important players in regulating various aspects of Streptomyces biology. Their exact roles in certain strain will ultimately depend on overall configuration of regulatory network and thus cannot be predicted on basis of in silico studies. Here, we explored S. ghanaensis gene SSFG_07725 (afsA) presumably involved in initial steps of formation of γ-butyrolactone LMWC. Disruption of afsA impaired aerial mycelium formation and increased the transcription of pleiotropic regulatory gene adpA, whereas level of moenomycin production remained virtually unaffected. We provide evidence that morphogenetic deficiency of afsA-minus mutant was caused by inability to produce diffusible LMWC. Possible links between γ-butyrolactone signaling and various aspects of S. ghanaensis biology are discussed.
Moenomycins (Mm) – phosphoglycolipid compounds produced by Streptomyces ghanaensis ATCC14672 – are considered a promising model for development of novel class of antibiotics. In this regard it is important to generate Mm overproducing strains which would be a basis for economically justified production of this antibiotic. In this work a set of genes for synthesis and reception of low-molecular weight signaling molecules (LSM) in ATCC14672 were described and their significance for Mm production was studied. The ATCC14672 genome carries structural and regulatory genes for production of LSMs of avenolide and γ-butyrolactone families. Additional copies of LSM biosynthetic genes ssfg_07848 and ssfg_07725 did not alter the Mm production level. ATCC14672 LSMs are not capable of restoring the sporulation of butyrolactone-nonproducing mutant of S. griseus. Likewise, while the heterologous host S. lividans 1326 produced Mm, its mutant M707 (deficient in the butyrolactone synthase gene scbA) did not. Thus, while the natural level of LSMs production by ATCC14672 does not limit Mm synthesis, the former is essential for the synthesis of moenomycins.
Actinobacteria of genus Streptomyces attract great interest of researchers. Their genomes encode cryptic gene clusters for as-yet-unknown antibiotics; heterologous expression of metagenomic libraries in model Streptomyces strains allow to discover new classes of compounds. However, it is crucial to understand the rules that govern codon usage in streptomycete genes, if we are to maximize the chances and level of expression of foreign genes in Streptomyces. In this study we addressed two questions related to codon usage in streptomycete genomes. First, we explored if there are patterns of dicodon usage in Streptomyces. Second, we searched for significant differences in patterns of codon substitution in different families of orthologous genes at different phylogenetic depth and degree of essentiality. To this end, we revealed several codon context rules, which are mainly associated with anomalous frequency of G/C downstream of C-ending codons. We developed a new bioinformatics tool, based on previously described bubble plot approach, allowing matrix-like visualization of codon substitution patterns in the dataset. Using this tool, we show that transcriptional factors of AdpA family carry significant fraction of nonsynonymous substitutions, although changes in its pattern for different actinobacterial orders (and as compared to Streptomycetales) do not follow simple rules.
Досліджено вплив CdCl 2 , Pb(NO 3 ) 2 , CuCl 2 та ZnCl 2 на активність каталази та супероксиддисмутази клітин Desulfovibrio desulfuricans Ya-11. Найвищу активність каталази (42,67×10 -2 мкмоль×хв -1 ×мг -1 білка) спостерігали за тривалої дії Pb(NO 3 ) 2 . За дії інших солей металів вона змінюється залежно від їх концентрації та тривалості росту. На основі результатів досліджень уперше побудовано ряд дії солей важких металів на активність каталази бактерій D. desulfuricans Ya-11: Pb(NO 3 ) 2 > CuCl 2 > CdCl 2 > ZnCl 2 . Найвищу активність супероксиддисмутази (61,52×10 2 мкмоль×хв -1 ×мг -1 білка) спостерігали за тривалого впливу ZnCl 2 . Інші досліджувані солі спричиняли зростання активності даного ферменту зі збільшенням концентрації солі металу. Уперше побудовано ряд дії солей важких металів на активність каталази бактерій D. desulfuricans Ya-11: ZnCl 2 > Pb(NO 3 ) 2 > CuCl 2 > CdCl 2 . И. В. Кушкевич, C. А. Гнатуш, Г. В. Мутенко Львовский национальный университет им. Ивана Франко АКТИВНОСТЬ КАТАЛАЗЫ И СУПЕРОКСИДДИСМУТАЗЫ БАКТЕРИЙ DESULFOVIBRIO DESULFURICANS YA-11 ПОД ВЛИЯНИЕМ СОЛЕЙ ТЯЖЕЛЫХ МЕТАЛЛОВ Исследовано влияние CdCl 2 , Pb(NO 3 ) 2 , CuCl 2 и ZnCl 2 на активность каталазы и супероксиддисмутазы клеток Desulfovibrio desulfuricans Ya-11. Самую высокую активность каталазы (42,67×10 -2 мкмоль×мин. -1 ×мг -1 белка) определили при длительном действии Pb(NO 3 ) 2 . При влиянии солей других металлов она изменяется в зависимости от их концентрации и длительности роста. На основании результатов исследований впервые определен ряд действия солей тяжелых металлов на активность каталазы бактерий D. desulfuricans Ya-11: Pb(NO 3 ) 2 > CuCl 2 > CdCl 2 > ZnCl 2 . Самая высокая активность супероксиддисмутазы (61,52×10 2 мкмоль×мин. -1 ×мг -1 белка) определена при длительном влиянии ZnCl 2 . Соли других тяжелых металлов вызывали увеличение активности исследуемого фермента с возрастанием их концентраций. Впервые определен ряд действия солей тяжелых металлов на активность каталазы бактерий D. desulfuricans Ya-11: ZnCl 2 > Pb(NO 3 ) 2 > CuCl 2 > CdCl 2 .The highest catalase activity (42.67×10 -2 µM×min -1 ×mg -1 of protein) in the cells of Desulfovibrio desulfuricans Ya-11 has been observed under the prolonged Pb(NO 3 ) 2 influence. In the presence of other heavy metals' salts it has been changed in dependence on their concentrations and growth duration. Based on our research data the row of heavy metals' salts by its influence on D. desulfuricans Ya-11 catalase acti-5©
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