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Erythromycin‐induced ribosome stalling and RNase J1‐mediated mRNA processing in Bacillus subtilis
Author(s) -
Yao Shiyi,
Blaustein Joshua B.,
Bechhofer David H.
Publication year - 2008
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2008.06370.x
Subject(s) - endoribonuclease , biology , shine dalgarno sequence , ribosomal binding site , ribosome , rnase p , start codon , coding region , messenger rna , translational frameshift , transfer rna , microbiology and biotechnology , bacillus subtilis , genetics , rna , gene , bacteria
Summary 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.