Recruitment of sigma54-RNA polymerase to the Pu promoter of Pseudomonas putida through integration host factor-mediated positioning switch of alpha subunit carboxyl-terminal domain on an UP-like element

The Journal of Biological Chemistry
Raffaella MacchiG Bertoni

Abstract

The interactions between the sigma54-containing RNA polymerase (sigma54-RNAP) and the region of the Pseudomonas putida Pu promoter spanning from the enhancer to the binding site for the integration host factor (IHF) were analyzed both by DNase I and hydroxyl radical footprinting. A short Pu region centered at position -104 was found to be involved in the interaction with sigma54-RNAP, both in the absence and in the presence of IHF protein. Deletion or scrambling of the -104 region strongly reduced promoter affinity in vitro and promoter activity in vivo, respectively. The reduction in promoter affinity coincided with the loss of IHF-mediated recruitment of the sigma54-RNAP in vitro. The experiments with oriented-alpha sigma54-RNAP derivatives containing bound chemical nuclease revealed interchangeable positioning of only one of the two alpha subunit carboxyl-terminal domains (alphaCTDs) both at the -104 region and in the surroundings of position -78. The addition of IHF resulted in perfect position symmetry of the two alphaCTDs. These results indicate that, in the absence of IHF, the sigma54-RNAP asymmetrically uses only one alphaCTD subunit to establish productive contacts with upstream sequences of the Pu promoter. In the pre...Continue Reading

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Citations

Feb 3, 2005·Nature Reviews. Microbiology·Ildefonso Cases, Víctor de Lorenzo
Feb 9, 2010·Nature Reviews. Microbiology·Shane C Dillon, Charles J Dorman
Apr 28, 2009·Nucleic Acids Research·Yi-Xin HuoYi-Ping Wang
Aug 3, 2012·Molecular Plant-microbe Interactions : MPMI·Christina RamelRobert Dudler
Jun 5, 2007·Biophysical Chemistry·Chiara ZurlaDavid D Dunlap
Nov 9, 2010·FEMS Microbiology Reviews·Victoria Shingler
Jun 22, 2010·FEMS Microbiology Reviews·Montserrat Elías-ArnanzFrancisco J Murillo
Jan 29, 2019·Frontiers in Microbiology·Ritika ChatterjeeDipshikha Chakravortty
Feb 6, 2020·Journal of Basic Microbiology·Tingting HuangMorigen Moregen
Nov 15, 2008·The Journal of Biological Chemistry·Lisandro M D BernardoVictoria Shingler

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