Supportive and inhibitory elements of a putative PrfA‐dependent promoter in Listeria monocytogenes

• Published in: Molecular microbiology 2005-02, Vol.55 (4), p.986-997
• Main Authors: Luo, Qin, Herler, Michael, Müller‐Altrock, Stefanie, Goebel, Werner
• Format: Article
• Language: English
• Subjects: 3-Phosphoshikimate 1-Carboxyvinyltransferase; Alkyl and Aryl Transferases - genetics; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Base Sequence; Binding sites; Biological and medical sciences; DNA Primers; DNA-Directed RNA Polymerases - metabolism; Fundamental and applied biological sciences. Psychology; Genes; Genetics; Listeria monocytogenes; Listeria monocytogenes - genetics; Microbiology; Miscellaneous; Models, Molecular; Molecular Sequence Data; Nucleic Acid Conformation; Peptide Termination Factors; Promoter Regions, Genetic - genetics; RNA polymerase; Sequence Alignment; Sequence Homology, Nucleic Acid; Trans-Activators - genetics; Trans-Activators - metabolism; Transcription, Genetic
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/j.1365-2958.2005.04417.x

Summary Elements essential for PrfA‐dependent transcription were analysed on two promoters of Listeria monocytogenes, the PrfA‐dependent promoter of the phospholipase gene plcA (PplcA) and a putative promoter of the aroA gene (ParoA2) which contains a similar PrfA‐binding site and a similar −10 box as PplcA but does not function as PrfA‐dependent promoter. We constructed a series of hybrid plcA‐aroA promoters by exchanging corresponding sequence elements of these two ‘promoters’. The results showed that the two critical elements of PrfA‐dependent promoters, the PrfA‐box and the −10 box, can be functionally exchanged as long as the distance in between is maintained to 22 or 23 bp. However, the interspace sequence and the sequence downstream of the −10 box of ParoA2 were strongly inhibitory for PrfA‐dependent transcription. A detailed analysis of these two sequences revealed that the RNA polymerase binding site being part of the actual in vivo and in vitro used aroA promoter (ParoA1) and a sequence immediately downstream of the putative −10 site, possibly blocking the formation of the open complex, were responsible for the inhibition of PrfA‐dependent transcription from ParoA2. Taking into consideration the lessons learned from this study we were able to construct a functional PrfA‐dependent aroA promoter.