Differential interaction of the transcription factor PrfA and the PrfA-activating factor (Paf) of Listeria monocytogenes with target sequences

Molecular Microbiology
C DickneiteZ Sokolovic

Abstract

The interaction of the purified PrfA transcription factor with the regulatory sequences located upstream of the PrfA-dependent listeriolysin (hly) and internalin (inlA) genes was studied in the presence and in the absence of Paf (PrfA-activating factor)-containing extracts. It is shown that PrfA protein is able to bind, independently of additional factors, to a 109bp DNA fragment including the entire hly promoter sequence with the anticipated PrfA binding site ('PrfA-box'). PrfA alone, but not in combination with Paf, can also bind to a shorter target sequence of 28 bp comprising essentially the PrfA-box of the hly promoter. The addition of a Paf-containing extract does not lead to significant protein binding to these two hly target sequences in the absence of PrfA but converts the complex (CIII) consisting of PrfA and the 109 bp hly DNA fragment to a slower migrating PrfA-Paf-DNA complex (CI). Incubation of cell-free extracts of wild-type Listeria monocytogenes with the 109 bp DNA fragment leads to the formation of CI. The addition of polyclonal PrfA antibodies causes a supershift of CIII. Purified PrfA and PrfA-Paf also bind to a DNA fragment containing the PrfA-dependent promoter P2 of inlA, albeit at a lower rate when compa...Continue Reading

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Citations

Oct 12, 2000·International Journal of Food Microbiology·M Sheikh-ZeinoddinC E Rees
Jan 16, 2007·Applied and Environmental Microbiology·Dmitriy V VolokhovAnthony D Hitchins
Feb 15, 2011·Applied and Environmental Microbiology·Peter RossmanithMartin Wagner
Jul 4, 2001·Clinical Microbiology Reviews·J A Vázquez-BolandJ Kreft
Feb 21, 2002·Infection and Immunity·Lynne M Shetron-RamaNancy E Freitag
Sep 19, 2006·Journal of Bacteriology·Norman MauderStefanie Müller-Altrock
Sep 19, 2006·Journal of Bacteriology·Christine RotterBirgit Scharf
Jul 5, 2001·International Journal of Medical Microbiology : IJMM·J Kreft, J A Vázquez-Boland
Oct 27, 2001·Molecular Microbiology·M Lalic-MülthalerW Goebel

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