Fig. 8: RNase Y-dependent pilA2 mRNA cleavage is necessary for the temperature-dependent regulation of sipW expression. | npj Biofilms and Microbiomes

Fig. 8: RNase Y-dependent pilA2 mRNA cleavage is necessary for the temperature-dependent regulation of sipW expression.

From: Temperature-regulated heterogeneous extracellular matrix gene expression defines biofilm morphology in Clostridium perfringens

Fig. 8

a Schematics of the posttranscriptional regulation model of the pilA2 gene. We constructed a deletion mutant of the 5′ UTR of pilA2, which contains the cleavage site. b Nucleotide sequence of the upstream region of the pilA2 gene. A triangle indicates a posttranscriptional cleavage site. Inverted arrows represent an inverted repeat sequence. The predicted ΔG value of the stem-loop structure is −4.60 kcal/mol. c Northern and western blotting related to pilA2. Cells were cultured to mid-exponential phase (OD600 = 1.0) at 37 °C. The 23S rRNAs stained with methylene blue are shown at the bottom as an RNA loading control. As a protein loading control, the protein that nonspecifically reacted with the antibodies are shown at the bottom. d Quantitative analysis of the sipW promoter in the pilA2 5′ UTR deletion mutant by FACS analysis. We analyzed 100,000 events for each sample. e Quantitative analysis of the sipW promoter in the RNase Y-depleted strain by FACS analysis. We analyzed 100,000 events for each sample.

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