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The stringent response does not influence ribosome pausing in Bacillus subtilis

Title: The stringent response does not influence ribosome pausing in Bacillus subtilis
Authors: Han, Yaozu; Agnolin, Alberto; van der Kloet, Frans; Sauer, Christopher; Costea, Paul Igor; Felle, Max Fabian; Appelbaum, Mathis; Hamoen, Leendert W
Contributors: China Scholarship Council; BASF
Source: Nucleic Acids Research ; volume 53, issue 20 ; ISSN 0305-1048 1362-4962
Publisher Information: Oxford University Press (OUP)
Publication Year: 2025
Description: The stringent response represses translation and is activated when cells enter the stationary phase and intracellular amino acid levels drop. Bacillus subtilis, a well-known industrial enzyme production organism, secretes most enzymes during the stationary phase. However, translation pausing can affect protein folding and production. Here, we examined whether the stringent response affects ribosome pausing in B. subtilis. To investigate this, genome-wide ribosome profiling was performed using a stringent response mutant that overexpressed the α-amylase AmyM. Blocking the stringent response did increase overall protein synthesis, however AmyM production was reduced. Ribosome profiling revealed that this was not caused by a reduction in translation. In fact, absence of the stringent response did not markedly influence ribosome pausing. Late into the stationary phase an increased ribosome pausing at tryptophan codons was observed, suggesting a depletion of tryptophan. A strong suppression of tryptophan biosynthesis and acquisition genes, under control of the trp RNA-binding attenuation protein TRAP, likely accounts for this. Why this occurs is unclear since TRAP does not belong to the stringent response regulon of B. subtilis. Finally, the genome-wide ribosome profiles revealed several operon genes with unusually low translation activities, illustrating the importance of translation initiation as a regulatory element of expression.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1093/nar/gkaf1088
Availability: https://doi.org/10.1093/nar/gkaf1088; https://academic.oup.com/nar/article-pdf/53/20/gkaf1088/65048724/gkaf1088.pdf
Rights: https://creativecommons.org/licenses/by-nc/4.0/
Accession Number: edsbas.1C1E620F
Database: BASE