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J. Biol. Chem., Vol. 276, Issue 11, 8159-8164, March 16, 2001
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From the The bacterial twin arginine translocation (Tat)
pathway is capable of exporting cofactor-containing enzymes into the
periplasm. To assess the capacity of the Tat pathway to export
heterologous proteins and to gain information about the property of the
periplasm, we fused the twin arginine signal peptide of the
trimethylamine N-oxide reductase to the jellyfish green
fluorescent protein (GFP). Unlike the Sec pathway, the Tat system
successfully exported correctly folded GFP into the periplasm of
Escherichia coli. Interestingly, GFP appeared as a halo in
most cells and occasionally showed a polar localization in wild type
strains. When subjected to a mild osmotic up-shock, GFP relocalized
very quickly at the two poles of the cells. The conversion from the
halo structure to a periplasmic gathering at particular locations was
also observed with spherical cells of the
Translocation of Jellyfish Green Fluorescent Protein via the
Tat System of Escherichia coli and Change of Its
Periplasmic Localization in Response to Osmotic Up-shock*
,,,,
Laboratoire de Chimie Bactérienne,
UPR9043 and the § Service de Microscopie Electronique,
Institut de Biologie Structurale et Microbiologie, CNRS, 31 chemin
Joseph Aiguier, F-13402 Marseille cedex 20 and the ¶ Equipe
Osmoadaptation chez les Bactéries, Université de Rennes 1,
Campus de Beaulieu, UMR CNRS 6026, F-35042 Rennes, France
rodA-pbpA mutant or of the wild type strain
treated with lysozyme. Therefore, the periplasm is not a uniform
compartment and the polarization of GFP is unlikely to be caused
by simple invagination of the cytoplasmic membrane at the poles.
Moreover, the polar gathering of GFP is reversible; the reversion was
accelerated by glucose and inhibited by azide and carbonyl cyanide
m-chlorophenylhydrazone, indicating an active adaptation of
the bacteria to the osmolarity in the medium. These results strongly
suggest a relocalization of periplasmic substances in response to
environmental changes. The polar area might be the preferential zone
where bacteria sense the change in the environment.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by "Quality of Life and Management of Living
Resources" Grant QLK3-CT-1999. To whom correspondence should be
addressed. Tel.: 33-491164157; Fax: 33-491718914; E-mail:
wu@ibsm.cnrs-mrs.fr.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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