RESUMO
Ms1 is a sRNA recently found in mycobacteria and several other actinobacterial species. Ms1 interacts with the RNA polymerase (RNAP) core devoid of sigma factors, which differs from 6S RNA that binds to RNAP holoenzymes containing the primary sigma factor. Here we show that Ms1 is the most abundant non-rRNA transcript in stationary phase in Mycobacterium smegmatis. The accumulation of Ms1 stems from its high-level synthesis combined with decreased degradation. We identify the Ms1 promoter, PMs1 , and cis-acting elements important for its activity. Furthermore, we demonstrate that PNPase (an RNase) contributes to the differential accumulation of Ms1 during growth. Then, by comparing the transcriptomes of wt and ΔMs1 strains from stationary phase, we reveal that Ms1 affects the intracellular level of RNAP. The absence of Ms1 results in decreased levels of the mRNAs encoding ß and ß' subunits of RNAP, which is also reflected at the protein level. Thus, the ΔMs1 strain has a smaller pool of RNAPs available when the transcriptional demand increases. This contributes to the inability of the ΔMs1 strain to rapidly react to environmental changes during outgrowth from stationary phase.
Assuntos
RNA Polimerases Dirigidas por DNA/metabolismo , Mycobacterium smegmatis/enzimologia , Mycobacterium smegmatis/metabolismo , RNA Bacteriano/metabolismo , Pequeno RNA não Traduzido/metabolismo , Deleção de Genes , Perfilação da Expressão Gênica , Mycobacterium smegmatis/genética , Mycobacterium smegmatis/crescimento & desenvolvimento , Pequeno RNA não Traduzido/genéticaRESUMO
The anthocyanin composition of blue (Triticum aestivum L., cv. Skorpion) and purple wheat (Triticum aethiopicum JAKUBZ cv. Abyssinskaja arrasajta cv. Abyssinskaja arrasajta), cultivated in the Czech Republic, and of the prepared whole blue and purple wheat bread was determined. In blue and purple wheat, 19 and 26 anthocyanins, respectively, were tentatively identified by liquid chromatography and mass spectrometry. The total content of anthocyanins determined in blue and purple wheat was 9.26 and 13.23 mgkg(-1), respectively. The breads were baked at 240 and 180 °C. Some significant differences in anthocyanins content were observed between breads prepared at different baking temperatures. The content of cyanidin-3-glucoside, delphinidin-3-glucoside and pelargonidin-3-glucoside was determinated in starting material, whole meal flours and baked breads. These kinds of wheat are suitable for baking bread, since intake of anthocyanins may play an important role in the prevention of human diseases.