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PMID: 7533884 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The mRNA for the 23S rRNA methylase encoded by the ermE gene of Saccharopolyspora erythraea is translated in the absence of a conventional ribosome-binding site.

Molecular microbiology ·Vol. 14 ·No. 3 ·1994-11-00 ·Pages 533-45

Bibb MJ, White J, Ward JM, Janssen GR

Abstract

Transcriptional analysis of the ermE gene of Saccharopolyspora erythraea, which confers resistance to erythromycin by N6-dimethylation of 23S rRNA and which is expressed from two promoters, ermEp1 and ermEp2, revealed a complex regulatory region in which transcription is initiated in a divergent and overlapping manner. Two promoters (eryC1p1 and eryC1p2) were identified for the divergently transcribed erythromycin biosynthetic gene eryC1, which plays a role in the formation of desosamine or its attachment to the macrolide ring. Transcription from eryC1p2 starts at the same position as that of ermEp1, but on the opposite strand of the DNA helix, suggesting co-ordinate regulation of genes for erythromycin production and resistance. ermEp1 initiates transcription at, and one nucleotide before, the ermE translational start codon. Site-directed and deletion mutagenesis, combined with immunochemical analysis, demonstrated that the ermEp1 transcript is translated in the absence of a conventional ribosome-binding site to give rise to the full-length 23S rRNA methylase. Deletion of the -35 region of ermEp1 reduced, but did not abolish, promoter activity, reminiscent of the 'extended -10' class of bacterial promoters which, like ermEp1, possess TGN motifs immediately upstream of their -10 regions and which initiate transcription seven nucleotides downstream of the -10 region.

Related Genes
MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Codon/genetics DNA, Bacterial/genetics Drug Resistance, Microbial/genetics Erythromycin/pharmacology Genes, Bacterial Methyltransferases/genetics Molecular Sequence Data Mutagenesis, Site-Directed Promoter Regions, Genetic Protein Biosynthesis RNA, Bacterial/genetics RNA, Messenger/genetics Restriction Mapping Ribosomes/metabolism Saccharopolyspora/drug effects,enzymology,genetics Sequence Deletion Sequence Homology, Nucleic Acid Transcription, Genetic
Chemicals
Codon DNA, Bacterial RNA, Bacterial RNA, Messenger Erythromycin Methyltransferases rRNA (adenosine-O-2'-)methyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bibb M J
John Innes Centre, Colney, Norwich, UK.
White J
Ward J M
Janssen G R
Article Info
Journal
Molecular microbiology
Abbr.
Mol Microbiol
ISSN
0950-382X
Published
1994-11-00
Pages
533-45
Language
English
Region
England
NLM ID
8712028
Subset
IM
Databases
GENBANK
M11200
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