Identification of Ruminiclostridium cellulolyticum endoribonuclease specific to cip-cel mRNA

Abstract:

[Objective] In this study, the endoribonuclease processing thecip-cel mRNA encoding cellulosome was identified inRuminiclostridium cellulolyticum. [Methods] The activity of four putative endoribonucleases (RNase III, RNase J, RNase G, and RNase Y) in cip-cel mRNA cleavage was analyzed via gene knockout by Clostron, overexpression in vivo, overexpression in vitro, and activity analysis. [Results]Genes (rnc and rnj) encoding RNase III and RNase J were disrupted and the resulted mutants did not affect the processing in intergenic region (IR) of the cip-cel mRNA. Moreover, RNase G and RNase Y were overexpressed and purified in vitro. RNase Y could cleave and degrade the mRNA harboring IR of the cip-cel mRNA in vitro, while RNase G failed to have any effect on that. Furthermore, overexpression of RNase Y in vivo could accelerate the degradation of cip-cel mRNA. [Conclusion]The cip-cel mRNA is potentially processed by RNase Y. The result helps deepen the understanding of the function of RNase Y in Gram-positive bacteria and the enzyme’s regulation of differential gene expression at the post-transcription level.