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- Title
Erythromycin-induced ribosome stalling and RNase J1-mediated mRNA processing in Bacillus subtilis.
- Authors
Shiyi Yao; Blaustein, Joshua B.; Bechhofer, David H.
- Abstract
Addition of erythromycin (Em) to a Bacillus subtilis strain carrying the ermC gene results in ribosome stalling in the ermC leader peptide coding sequence. Using Δ ermC, a deletion derivative of ermC that specifies the 254 nucleotide Δ ermC mRNA, we showed previously that ribosome stalling is concomitant with processing of Δ ermC mRNA, generating a 209 nucleotide RNA whose 5′ end maps to codon 5 of the Δ ermC coding sequence. Here we probed for peptidyl-tRNA to show that ribosome stalling occurs after incorporation of the amino acid specified by codon 9. Thus, cleavage upstream of codon 5 is not an example of ‘A-site cleavage’ that has been reported for Escherichia coli. Analysis of Δ ermC mRNA processing in endoribonuclease mutant strains showed that this processing is RNase J1-dependent. Δ ermC mRNA processing was inhibited by the presence of stable secondary structure at the 5′ end, demonstrating 5′-end dependence, and was shown to be a result of RNase J1 endonuclease activity, rather than 5′-to-3′ exonuclease activity. Examination of processing in derivatives of Δ ermC that had codons inserted upstream of the ribosome stalling site revealed that Em-induced ribosome stalling can occur considerably further from the start codon than would be expected based on previous studies.
- Publication
Molecular Microbiology, 2008, Vol 69, Issue 6, p1439
- ISSN
0950-382X
- Publication type
Academic Journal
- DOI
10.1111/j.1365-2958.2008.06370.x