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Journal of Bacteriology, July 2009, p. 4410-4418, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.00215-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Subcellular Localization of TatAd of Bacillus subtilis Depends on the Presence of TatCd or TatCy{triangledown}

Anja N. J. A. Ridder, Esther J. de Jong,{dagger} Jan D. H. Jongbloed,{dagger} and Oscar P. Kuipers*

Department of Genetics, University of Groningen, Groningen Biomolecular Sciences and Biotechnology Institute, Kerklaan 30, 9751 NN Haren, The Netherlands

Received 18 February 2009/ Accepted 17 April 2009

The gram-positive bacterium Bacillus subtilis contains two minimal Tat translocases, TatAdCd and TatAyCy, which are each involved in the secretion of one or more specific protein substrates. We have investigated the subcellular localization of the TatA components by employing C-terminal green fluorescent protein (GFP) fusions and fluorescence microscopy. When expressed from a xylose-inducible promoter, the TatA-GFP fusion proteins displayed a dual localization pattern, being localized peripherally and showing bright foci which are predominantly located at the division sites and/or poles of the cells. Importantly, the localization of TatAd-GFP was similar when the protein was expressed from its own promoter under phosphate starvation conditions, indicating that these foci are not the result of artificial overexpression. Moreover, the TatAd-GFP fusion protein was shown to be functional in the translocation of its substrate PhoD, provided that TatCd is also present. Furthermore, we demonstrate that the localization of TatAd-GFP in foci depends on the presence of the TatCd component. Remarkably, however, the TatAd-GFP foci can also be observed in the presence of TatCy, indicating that TatAd can interact not only with TatCd but also with TatCy. These results suggest that the formation of TatAd complexes in B. subtilis is controlled by TatC.


* Corresponding author. Mailing address: Department of Genetics, University of Groningen, Groningen Biomolecular Sciences and Biotechnology Institute, Kerklaan 30, 9751 NN Haren, The Netherlands. Phone: (31) 50 363 2093. Fax: (31) 50 363 2348. E-mail: o.p.kuipers{at}rug.nl

{triangledown} Published ahead of print on 24 April 2009.

{dagger} Present address: Genetica UMCG, Postbus 30001, 9700 RB Groningen, The Netherlands.


Journal of Bacteriology, July 2009, p. 4410-4418, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.00215-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.