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The role of the sulfur globule proteins of Allochromatium vinosum: mutagenesis of the sulfur globule protein genes and expression studies by real-time RT-PCR

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Abstract

During oxidation of reduced sulfur compounds, the purple sulfur bacterium Allochromatium vinosum stores sulfur in the periplasm in the form of intracellular sulfur globules. The sulfur in the globules is enclosed by a protein envelope that consists of the homologous 10.5-kDa proteins SgpA and SgpB and the smaller 8.5-kDa SgpC. Reporter gene fusions of sgpA and alkaline phosphatase showed the constitutive expression of sgpA in A. vinosum and yielded additional evidence for the periplasmic localization of the sulfur globules. Expression analysis of the wild-type sgp genes by quantitative RT-PCR using the LightCycler system showed the constitutive expression of all three sgp genes. The expression of sgpB and sgpC is significantly enhanced under photolithotrophic conditions. Interestingly, sgpB is expressed ten times less than sgpA and sgpC implying that SgpA and SgpC are the “main proteins” of the sulfur globule envelope. Mutants with inactivated sgpA or sgpB did not show any differences in comparison with the wild-type, i.e., the encoded proteins can replace each other, whereas inactivation of sgpC leads to the formation of considerably smaller sulfur globules. This indicates a role of SgpC for globule expansion. A sgpBC double mutant was unable to grow on sulfide and could not form sulfur globules, showing that the protein envelope is indispensible for the formation and deposition of intracellular sulfur.

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Abbreviations

Sgp:

Sulfur globule protein

sgp :

Sulfur globule protein gene

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Acknowledgments

We thank Daniel C Brune (Arizona State University, Tempe, USA) for useful hints concerning HPLC analysis of the sulfur globule proteins, Brigitte Kühlmorgen for skilfull technical assistance with electron microscopy, and Kobchai Pattaragulwanit (Chulalongkorn University, Bangkok, Thailand) for helpful discussions. AP thanks the GEN-IAL GmbH (Troisdorf) for the possibility to perform the LightCycler experiments in their laboratory and the Stiftung der Deutschen Wirtschaft (Studienförderwerk Klaus Murmann) for a doctoral scholarship. This work was supported by the Deutsche Forschungsgemeinschaft (DFG grant Tr 133/26-1,2,3) and the Fonds der Chemischen Industrie (HGT).

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  1. Alexander Prange

    Present address: Institut für Pflanzenkrankheiten, Abteilung Landwirtschaftliche und Lebensmittel-Mikrobiologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Meckenheimer Allee 168, 53115, Bonn, Germany

Authors and Affiliations

  1. Institut für Mikrobiologie & Biotechnologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Meckenheimer Allee 168, 53115, Bonn, Germany

    Alexander Prange, Hans G. Trüper & Christiane Dahl

  2. IBELIN GbR, Nordring 15, 59929, Brilon, Germany

    Alexander Prange

  3. Max-Planck-Institut für Biochemie, Am Klopferspitz 18, 82152, Martinsried, Germany

    Harald Engelhardt

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  1. Alexander Prange
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Correspondence to Christiane Dahl.

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The paper is dedicated to Prof. Dr. Dr. h.c. mult. Hans Günter Schlegel, Göttingen, on the occasion of his 80th birthday on October 24th, 2004, with great gratitude, as our interest in microbial sulfur metabolism goes back to the early 1960s, when HGT worked in Prof. Schlegels laboratory and in 1972 established this field in Bonn.

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Prange, A., Engelhardt, H., Trüper, H.G. et al. The role of the sulfur globule proteins of Allochromatium vinosum: mutagenesis of the sulfur globule protein genes and expression studies by real-time RT-PCR. Arch Microbiol 182, 165–174 (2004). https://doi.org/10.1007/s00203-004-0683-3

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