期刊
MOLECULAR MICROBIOLOGY
卷 69, 期 6, 页码 1439-1449出版社
WILEY-BLACKWELL
DOI: 10.1111/j.1365-2958.2008.06370.x
关键词
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资金
- National Institutes of Health [GM-48804]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM048804] Funding Source: NIH RePORTER
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 Delta ermC, a deletion derivative of ermC that specifies the 254 nucleotide Delta ermC mRNA, we showed previously that ribosome stalling is concomitant with processing of Delta ermC mRNA, generating a 209 nucleotide RNA whose 5' end maps to codon 5 of the Delta 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 Delta ermC mRNA processing in endoribonuclease mutant strains showed that this processing is RNase J1-dependent. Delta 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 Delta 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.
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