Effect of the deletion of qmoABC and the promoter distal gene encoding a hypothetical protein on sulfate-reduction in Desulfovibrio vulgaris Hildenborough

The pathway of electrons required for the reduction of sulfate in sulfate-reducing bacteria (SRB) is not yet fully characterized. In order to determine the role of a transmembrane protein complex suggested to be involved in this process, a deletion of Desulfovibrio vulgaris Hildenborough was created by marker exchange mutagenesis that eliminated four genes putatively encoding the QmoABC complex and a hypothetical protein (DVU0851). The Qmo complex (quinone-interacting membrane-bound oxidoreductase) is proposed to be responsible for transporting electrons to the dissimilatory adenosine-5?phosphosulfate (APS) reductase in SRB. In support of the predicted role of this complex, the deletion mutant was unable to grow using sulfate as its sole electron acceptor with a range of electron donors. To explore a possible role for the hypothetical protein in sulfate reduction, a second mutant was constructed that had lost only the gene that codes for DVU0851. The second constructed mutant grew with sulfate as the sole electron acceptor; however, there was a lag that was not present with the wild-type or complemented strain. Neither deletion strain was significantly impaired for growth with sulfite or thiosulfate as terminal electron acceptor. Complementation of the D(qmoABC-DVU0851) mutant with all four genes or only the qmoABC genes restored its ability to grow by sulfate respiration. These results confirmed the prediction that the Qmo complex is in the electron pathway for sulfate-reduction and revealed that no other transmembrane complex could compensate when Qmo was lacking.

[1]  S. Ho,et al.  Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. , 2013, BioTechniques.

[2]  J. Wall,et al.  Development of a Markerless Genetic Exchange System for Desulfovibrio vulgaris Hildenborough and Its Use in Generating a Strain with Increased Transformation Efficiency , 2009, Applied and Environmental Microbiology.

[3]  Jizhong Zhou,et al.  Energy metabolism in Desulfovibrio vulgaris Hildenborough: insights from transcriptome analysis , 2008, Antonie van Leeuwenhoek.

[4]  J. Kuever,et al.  Homology Modeling of Dissimilatory APS Reductases (AprBA) of Sulfur-Oxidizing and Sulfate-Reducing Prokaryotes , 2008, PloS one.

[5]  S. Elledge,et al.  Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC , 2007, Nature Methods.

[6]  R. Amann,et al.  Clustered Genes Related to Sulfate Respiration in Uncultured Prokaryotes Support the Theory of Their Concomitant Horizontal Transfer , 2005, Journal of bacteriology.

[7]  E. A. Greene,et al.  Gene expression analysis of the mechanism of inhibition of Desulfovibrio vulgaris Hildenborough by nitrate-reducing, sulfide-oxidizing bacteria. , 2005, Environmental microbiology.

[8]  Inna Dubchak,et al.  Reconstruction of regulatory and metabolic pathways in metal-reducing δ-proteobacteria , 2004, Genome Biology.

[9]  Rekha Seshadri,et al.  The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough , 2004, Nature Biotechnology.

[10]  M. Teixeira,et al.  A novel membrane-bound respiratory complex from Desulfovibrio desulfuricans ATCC 27774. , 2003, Biochimica et biophysica acta.

[11]  J. Heidelberg,et al.  Gene Expression Analysis of Energy Metabolism Mutants of Desulfovibrio vulgaris Hildenborough Indicates an Important Role for Alcohol Dehydrogenase , 2003, Journal of bacteriology.

[12]  Ronald W. Davis,et al.  Functional profiling of the Saccharomyces cerevisiae genome , 2002, Nature.

[13]  L. Krumholz,et al.  Desulfovibrio sp. Genes Involved in the Respiration of Sulfate during Metabolism of Hydrogen and Lactate , 2002, Applied and Environmental Microbiology.

[14]  G. Voordouw,et al.  Deletion of the hmc operon of Desulfovibrio vulgaris subsp. vulgaris Hildenborough hampers hydrogen metabolism and low-redox-potential niche establishment , 2000, Archives of Microbiology.

[15]  K. Struhl,et al.  Introduction of Plasmid DNA into Cells , 2000, Current protocols in neuroscience.

[16]  T. Brown Analysis of RNA by Northern and Slot‐Blot Hybridization , 1993, Current protocols in immunology.

[17]  G. Voordouw,et al.  Nucleotide sequence of dcrA, a Desulfovibrio vulgaris Hildenborough chemoreceptor gene, and its expression in Escherichia coli , 1992, Journal of bacteriology.

[18]  H. Cypionka,et al.  Sulfate formation via ATP sulfurylase in thiosulfate- and sulfite-disproportionating bacteria , 1989, Archives of Microbiology.

[19]  A. Roman,et al.  Alterations in the regulatory region of the human papillomavirus type 6 genome are generated during propagation in Escherichia coli , 1988, Journal of virology.

[20]  J. Wall,et al.  Properties of a hydrogen-inhibited mutant of Desulfovibrio desulfuricans ATCC 27774 , 1987, Journal of bacteriology.

[21]  R. Cord-Ruwisch A quick method for the determination of dissolved and precipitated sulfides in cultures of sulfate-reducing bacteria , 1985 .

[22]  H. D. Peck,et al.  Biochemistry of dissimilatory sulphate reduction. , 1982, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[23]  J. M. Odom,et al.  Hydrogen cycling as a general mechanism for energy coupling in the sulfate‐reducing bacteria, Desulfovibrio sp. , 1981 .

[24]  R. Thauer,et al.  Growth of Desulfovibrio species on Hydrogen and Sulphate as Sole Energy Source , 1981 .

[25]  Adam P. Arkin,et al.  Analysis of a ferric uptake regulator (Fur) mutant of Desulfovibrio vulgaris , 2010 .

[26]  Katherine H. Huang,et al.  The MicrobesOnline Web site for comparative genomics. , 2005, Genome research.