An unstructured 5′-coding region of the prfA mRNA is required for efficient translation
Open Access
- 2 November 2011
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 40 (4), 1818-1827
- https://doi.org/10.1093/nar/gkr850
Abstract
Expression of virulence factors in the human bacterial pathogen Listeria monocytogenes is almost exclusively regulated by the transcriptional activator PrfA. The translation of prfA is controlled by a thermosensor located in the 5′-untranslated RNA (UTR), and is high at 37°C and low at temperatures prfA -coding sequences were placed in front of lacZ . When expressed in Escherichia coli , the β-galactosidase expression was directly correlated to the length of the prfA -coding mRNA lying in front of lacZ . A similar effect was detected with gfp as a reporter gene in both L. monocytogenes and E. coli , emphasizing the requirement of the prfA- coding RNA for maximal expression. In vitro transcription/translation and mutational analysis suggests a role for the first 20 codons of the native prfA -mRNA for maximal expression. By toe-print and RNA-probing analysis, a flexible hairpin-loop located immediately downstream of the start-codon was shown to be important for ribosomal binding. The present work determines the importance of an unstructured part of the 5′-coding region of the prfA -mRNA for efficient translation.This publication has 30 references indexed in Scilit:
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