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Nucleic Acids Research Advance Access originally published online on December 1, 2007
Nucleic Acids Research 2008 36(2):598-606; doi:10.1093/nar/gkm1077
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Nucleic Acids Research, 2008, Vol. 36, No. 2 598-606
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Overlapping activator sequences determined for two oppositely oriented promoters in halophilic Archaea

Martina Bauer, Larissa Marschaus, Muriel Reuff, Verena Besche, Simone Sartorius-Neef and Felicitas Pfeifer*

Institut für Mikrobiologie und Genetik, TU Darmstadt, Schnittspahnstrasse 10, D-64287 Darmstadt, Germany

*To whom correspondence should be addressed. Tel: +06151 16 2957; Fax: +06151 16 2956; Email: pfeifer{at}bio.tu-darmstadt.de

Received August 31, 2007. Revised November 15, 2007. Accepted November 15, 2007.

Transcription of the genomic region involved in gas vesicle formation in Halobacterium salinarum (p-vac) and Haloferax mediterranei (mc-vac) is driven by two divergent promoters, PA and PD, separated by only 35 nt. Both promoters are activated by the transcription activator GvpE which in the case of PmcA requires a 20-nt sequence (UAS) consisting of two conserved 8-nt sequence portions located upstream of BRE. Here, we determined the two UAS elements in the promoter region of p-vac by scanning mutageneses using constructs containing PpD (without PpA) fused to the bgaH reporter gene encoding an enzyme with β-galactosidase activity, or the dual reporter construct pApD with PpD fused to bgaH and PpA to an altered version of gvpA. The two UAS elements found exhibited a similar extension and distance to BRE as previously determined for the UAS in PmcA. Their distal 8-nt portions almost completely overlapped in the centre of PpD–PpA, and mutations in this region negatively affected the GvpE-mediated activation of both promoters. Any alteration of the distance between BRE and UAS resulted in the loss of the GvpE activation, as did a complete substitution of the proximal 8-nt portion, underlining that a close location of UAS and BRE was very important.


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