Details
| Original language | English |
|---|---|
| Pages (from-to) | 82-88 |
| Number of pages | 7 |
| Journal | FEBS Letters |
| Volume | 569 |
| Issue number | 1-3 |
| Publication status | Published - 2 Jul 2004 |
Abstract
The Tat system has the ability to translocate folded proteins across the bacterial cytoplasmic membrane. In Escherichia coli, three functionally different translocon components have been identified, namely TatA, TatB, and TatC. These proteins were fused to the green fluorescent protein (GFP) and their localization was determined by confocal laser scanning fluorescence microscopy. TatA-GFP was distributed in the membrane, often with higher abundance at the poles. TatB-GFP was found in distinct spots at the poles of the cells. The fluorescence of TatC-GFP was very low and required a constitutive expression system to become higher than background, but then appearing polar. All three constructs complemented the chain-formation phenotype of corresponding mutant strains, indicating the functionality of the fusion proteins. TatB-GFP and TatC-GFP also complemented TMAO respiration deficiency and TatA-GFP the SDS-sensitivity of the mutant strains. The localization of the translocon-GFP fusions coincides with the fluorescence pattern of GFP fusions to Tat substrate signal sequences. We suggest that the active translocon complexes are mainly present at polar positions in Escherichia coli.
Keywords
- Fusion protein, GFP, green fluorescent protein, Green fluorescent protein, Protein transport, Tat, twin-arginine translocation, TatABC complex localization, TMAO, trimethylamine N-oxide, Twin-arginine translocation
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biophysics
- Biochemistry, Genetics and Molecular Biology(all)
- Structural Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: FEBS Letters, Vol. 569, No. 1-3, 02.07.2004, p. 82-88.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Localization of the Tat translocon components in Escherichia coli
AU - Berthelmann, Felix
AU - Brüser, Thomas
N1 - Funding Information: We thank Timothy Yahr and Matthias Müller for their generous donation of antisera. We are also very grateful to Tracy Palmer for sending us her tat deletion strains. We are indebted to Carsten Sanders and Tim Yahr for donation of plasmids. We thank Ute Lindenstrauss for excellent technical assistance, Anja Ebert for support with LSM techniques, Silke Trautmann for interest and fruitful communication, and Theresa Wermann for reading the manuscript. We especially thank Jan R. Andreesen for discussions and support. This work was supported by the Fonds der Chemischen Industrie and by the Deutsche Forschungsgemeinschaft (BR2285/1-1). Copyright: Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2004/7/2
Y1 - 2004/7/2
N2 - The Tat system has the ability to translocate folded proteins across the bacterial cytoplasmic membrane. In Escherichia coli, three functionally different translocon components have been identified, namely TatA, TatB, and TatC. These proteins were fused to the green fluorescent protein (GFP) and their localization was determined by confocal laser scanning fluorescence microscopy. TatA-GFP was distributed in the membrane, often with higher abundance at the poles. TatB-GFP was found in distinct spots at the poles of the cells. The fluorescence of TatC-GFP was very low and required a constitutive expression system to become higher than background, but then appearing polar. All three constructs complemented the chain-formation phenotype of corresponding mutant strains, indicating the functionality of the fusion proteins. TatB-GFP and TatC-GFP also complemented TMAO respiration deficiency and TatA-GFP the SDS-sensitivity of the mutant strains. The localization of the translocon-GFP fusions coincides with the fluorescence pattern of GFP fusions to Tat substrate signal sequences. We suggest that the active translocon complexes are mainly present at polar positions in Escherichia coli.
AB - The Tat system has the ability to translocate folded proteins across the bacterial cytoplasmic membrane. In Escherichia coli, three functionally different translocon components have been identified, namely TatA, TatB, and TatC. These proteins were fused to the green fluorescent protein (GFP) and their localization was determined by confocal laser scanning fluorescence microscopy. TatA-GFP was distributed in the membrane, often with higher abundance at the poles. TatB-GFP was found in distinct spots at the poles of the cells. The fluorescence of TatC-GFP was very low and required a constitutive expression system to become higher than background, but then appearing polar. All three constructs complemented the chain-formation phenotype of corresponding mutant strains, indicating the functionality of the fusion proteins. TatB-GFP and TatC-GFP also complemented TMAO respiration deficiency and TatA-GFP the SDS-sensitivity of the mutant strains. The localization of the translocon-GFP fusions coincides with the fluorescence pattern of GFP fusions to Tat substrate signal sequences. We suggest that the active translocon complexes are mainly present at polar positions in Escherichia coli.
KW - Fusion protein
KW - GFP, green fluorescent protein
KW - Green fluorescent protein
KW - Protein transport
KW - Tat, twin-arginine translocation
KW - TatABC complex localization
KW - TMAO, trimethylamine N-oxide
KW - Twin-arginine translocation
UR - http://www.scopus.com/inward/record.url?scp=3042589734&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2004.05.054
DO - 10.1016/j.febslet.2004.05.054
M3 - Article
C2 - 15225613
AN - SCOPUS:3042589734
VL - 569
SP - 82
EP - 88
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 1-3
ER -