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Translation of a minigene in the 5′ leader sequence of the enterohaemorrhagic Escherichia coli LEE1 transcription unit affects expression of the neighbouring downstream gene
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Islam, Md. Shahidul, Shaw, Robert K., Frankel, Gad, Pallen, Mark J. and Busby, Stephen J. W. (2012) Translation of a minigene in the 5′ leader sequence of the enterohaemorrhagic Escherichia coli LEE1 transcription unit affects expression of the neighbouring downstream gene. Biochemical Journal, 441 (1). pp. 247-253. doi:10.1042/BJ20110912 ISSN 0264-6021.
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Official URL: http://dx.doi.org/10.1042/BJ20110912
Abstract
The 5′ end of the major RNA transcript of the LEE1 operon of enterohaemorrhagic Escherichia coli contains ~170 bases before the AUG translation start codon of the first recognized gene, ler. This unusually long leader sequence carries three potential alternative AUG start codons. Using a lac fusion expression vector, we confirmed that the ler gene AUG is functional for translation initiation, and we checked for translation initiation at the three alternative AUG codons. Whereas two of the alternative AUG codons appear incompetent for translation initiation, we detected strong initiation at the third AUG, which is followed by one AAA codon and a UAG stop codon. The location of this very short two-codon open reading frame with respect to the ler translation start appears to be critical. Hence mutations that destroy the UAG stop codon, or short deletions between the UAG stop codon and the ler translation initiation region, result in big effects on ler expression. In the context of the full-length LEE1 operon leader sequence, translation of this very short two-codon open reading frame is necessary for optimal expression of the ler gene and for the subsequent interactions of enterohaemorrhagic Escherichia coli with host target cells.
Item Type: | Journal Article | ||||||||||
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Subjects: | Q Science > QR Microbiology | ||||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Microbiology & Infection Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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Library of Congress Subject Headings (LCSH): | RNA interference | ||||||||||
Journal or Publication Title: | Biochemical Journal | ||||||||||
Publisher: | Portland Press | ||||||||||
ISSN: | 0264-6021 | ||||||||||
Official Date: | 1 January 2012 | ||||||||||
Dates: |
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Volume: | 441 | ||||||||||
Number: | 1 | ||||||||||
Number of Pages: | 7 | ||||||||||
Page Range: | pp. 247-253 | ||||||||||
DOI: | 10.1042/BJ20110912 | ||||||||||
Status: | Peer Reviewed | ||||||||||
Publication Status: | Published | ||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||
Date of first compliant deposit: | 26 December 2015 | ||||||||||
Date of first compliant Open Access: | 26 December 2015 | ||||||||||
Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Wellcome Trust (London, England), Commonwealth Fund |
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