scholarly journals Novel Genes of the dsr Gene Cluster and Evidence for Close Interaction of Dsr Proteins during Sulfur Oxidation in the Phototrophic Sulfur Bacterium Allochromatium vinosum

2005 ◽  
Vol 187 (4) ◽  
pp. 1392-1404 ◽  
Author(s):  
Christiane Dahl ◽  
Sabine Engels ◽  
Andrea S. Pott-Sperling ◽  
Andrea Schulte ◽  
Johannes Sander ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Sulfur Oxidation ◽  
Integral Membrane Protein ◽  
Sulfite Reductase ◽  
Prosthetic Group ◽  
Allochromatium Vinosum ◽  
Sulfate Reducing ◽  
New Genes ◽  
Cytoplasmic Proteins ◽  
Sulfur Protein

ABSTRACT Seven new genes designated dsrLJOPNSR were identified immediately downstream of dsrABEFHCMK, completing the dsr gene cluster of the phototrophic sulfur bacterium Allochromatium vinosum D (DSM 180T). Interposon mutagenesis proved an essential role of the encoded proteins for the oxidation of intracellular sulfur, an obligate intermediate during the oxidation of sulfide and thiosulfate. While dsrR and dsrS encode cytoplasmic proteins of unknown function, the other genes encode a predicted NADPH:acceptor oxidoreductase (DsrL), a triheme c-type cytochrome (DsrJ), a periplasmic iron-sulfur protein (DsrO), and an integral membrane protein (DsrP). DsrN resembles cobyrinic acid a,c-diamide synthases and is probably involved in the biosynthesis of siro(heme)amide, the prosthetic group of the dsrAB-encoded sulfite reductase. The presence of most predicted Dsr proteins in A. vinosum was verified by Western blot analysis. With the exception of the constitutively present DsrC, the formation of Dsr gene products was greatly enhanced by sulfide. DsrEFH were purified from the soluble fraction and constitute a soluble α2β2γ2-structured 75-kDa holoprotein. DsrKJO were purified from membranes pointing at the presence of a transmembrane electron-transporting complex consisting of DsrKMJOP. In accordance with the suggestion that related complexes from dissimilatory sulfate reducers transfer electrons to sulfite reductase, the A. vinosum Dsr complex is copurified with sulfite reductase, DsrEFH, and DsrC. We therefore now have an ideal and unique possibility to study the interaction of sulfite reductase with other proteins and to clarify the long-standing problem of electron transport from and to sulfite reductase, not only in phototrophic bacteria but also in sulfate-reducing prokaryotes.

scholarly journals A Comparative Quantitative Proteomic Study Identifies New Proteins Relevant for Sulfur Oxidation in the Purple Sulfur Bacterium Allochromatium vinosum

2014 ◽  
Vol 80 (7) ◽  
pp. 2279-2292 ◽  
Author(s):  
Thomas Weissgerber ◽  
Marc Sylvester ◽  
Lena Kröninger ◽  
Christiane Dahl
Keyword(s):  
Sulfur Metabolism ◽  
Sulfur Oxidation ◽  
Hypothetical Protein ◽  
Mrna Levels ◽  
Reduced Sulfur Compounds ◽  
Allochromatium Vinosum ◽  
Purple Sulfur Bacterium ◽  
Content Type ◽  
Protein Levels ◽  
New Genes

ABSTRACTIn the present study, we compared the proteome response ofAllochromatium vinosumwhen growing photoautotrophically in the presence of sulfide, thiosulfate, and elemental sulfur with the proteome response when the organism was growing photoheterotrophically on malate. Applying tandem mass tag analysis as well as two-dimensional (2D) PAGE, we detected 1,955 of the 3,302 predicted proteins by identification of at least two peptides (59.2%) and quantified 1,848 of the identified proteins. Altered relative protein amounts (≥1.5-fold) were observed for 385 proteins, corresponding to 20.8% of the quantifiedA. vinosumproteome. A significant number of the proteins exhibiting strongly enhanced relative protein levels in the presence of reduced sulfur compounds are well documented essential players during oxidative sulfur metabolism, e.g., the dissimilatory sulfite reductase DsrAB. Changes in protein levels generally matched those observed for the respective relative mRNA levels in a previous study and allowed identification of new genes/proteins participating in oxidative sulfur metabolism. One gene cluster (hyd; Alvin_2036-Alvin_2040) and one hypothetical protein (Alvin_2107) exhibiting strong responses on both the transcriptome and proteome levels were chosen for gene inactivation and phenotypic analyses of the respective mutant strains, which verified the importance of the so-called Isp hydrogenase supercomplex for efficient oxidation of sulfide and a crucial role of Alvin_2107 for the oxidation of sulfur stored in sulfur globules to sulfite. In addition, we analyzed the sulfur globule proteome and identified a new sulfur globule protein (SgpD; Alvin_2515).

A comparison of the electrophoretic properties of the ATP-sulfurylases, APS-reductases, and sulfite reductases from cultures of dissimilatory sulfate-reducing bacteria

1973 ◽  
Vol 19 (3) ◽  
pp. 375-380 ◽  
Author(s):  
G. W. Skyring ◽  
P. A. Trudinger
Keyword(s):  
Desulfovibrio Desulfuricans ◽  
Sulfate Reducing Bacteria ◽  
Disc Electrophoresis ◽  
Sulfite Reductase ◽  
Electrophoretic Variation ◽  
Sulfate Reducing ◽  
Aps Reductase ◽  
Reducing Bacteria ◽  
Atp Sulfurylases

ATP-sulfurylases, APS-reductases, and sulfite reductases (SO3−2 → S−2) have been detected by gel disc electrophoresis in 13 cultures of dissimilatory sulfate-reducing bacteria and their electrophoretic properties have been compared. With respect to these three enzymes only, the results were indicative of some interspecies and intergenus homologies. In the Desulfovibrio strains (except Desulfovibrio desulfuricans 8301 which does not contain desulfoviridin), the major sulfite reductase was electrophoretically coincident with desulfoviridin and, in the Desulfotomaculum strains, with a brown protein. Some distinct patterns of electrophoretically distinguishable forms of APS-reductase were found. Considerable electrophoretic variation was found among the ATP-sulfurylases.

scholarly journals Identification of Two New Genes in thePseudomonasaeruginosaAmidase Operon, Encoding an ATPase (AmiB) and a Putative Integral Membrane Protein (AmiS)

1995 ◽  
Vol 270 (32) ◽  
pp. 18818-18824 ◽  
Author(s):  
Stuart A. Wilson ◽  
Rachel J. Williams ◽  
Laurence H. Pearl ◽  
Robert E. Drew
Keyword(s):  
Membrane Protein ◽  
Integral Membrane Protein ◽  
New Genes

scholarly journals Expansion of the Clavulanic Acid Gene Cluster: Identification and In Vivo Functional Analysis of Three New Genes Required for Biosynthesis of Clavulanic Acid by Streptomyces clavuligerus

2000 ◽  
Vol 182 (14) ◽  
pp. 4087-4095 ◽  
Author(s):  
Rongfeng Li ◽  
Nusrat Khaleeli ◽  
Craig A. Townsend
Keyword(s):  
Clavulanic Acid ◽  
Gene Cluster ◽  
Streptomyces Clavuligerus ◽  
Biosynthetic Gene Cluster ◽  
Gene Replacement ◽  
Resistant Bacteria ◽  
Acid Biosynthesis ◽  
Significant Similarity ◽  
New Genes

ABSTRACT Clavulanic acid is a potent inhibitor of β-lactamase enzymes and is of demonstrated value in the treatment of infections by β-lactam-resistant bacteria. Previously, it was thought that eight contiguous genes within the genome of the producing strainStreptomyces clavuligerus were sufficient for clavulanic acid biosynthesis, because they allowed production of the antibiotic in a heterologous host (K. A. Aidoo, A. S. Paradkar, D. C. Alexander, and S. E. Jensen, p. 219–236, In V. P. Gullo et al., ed., Development in industrial microbiology series, 1993). In contrast, we report the identification of three new genes, orf10 (cyp), orf11(fd), and orf12, that are required for clavulanic acid biosynthesis as indicated by gene replacement andtrans-complementation analysis in S. clavuligerus. These genes are contained within a 3.4-kb DNA fragment located directly downstream of orf9(cad) in the clavulanic acid cluster. While theorf10 (cyp) and orf11(fd) proteins show homologies to other knownCYP-150 cytochrome P-450 and [3Fe-4S] ferredoxin enzymes and may be responsible for an oxidative reaction late in the pathway, the protein encoded by orf12 shows no significant similarity to any known protein. The results of this study extend the biosynthetic gene cluster for clavulanic acid and attest to the importance of analyzing biosynthetic genes in the context of their natural host. Potential functional roles for these proteins are proposed.

Sirohydrochlorin. Prosthetic group of a sulfite reductase enzyme and its role in the biosynthesis of vitamin B12

1978 ◽  
Vol 100 (1) ◽  
pp. 316-318 ◽  
Author(s):  
A. I. Scott ◽  
A. J. Irwin ◽  
L. M. Siegel ◽  
J. N. Shoolery
Keyword(s):  
Vitamin B12 ◽  
Sulfite Reductase ◽  
Prosthetic Group

scholarly journals Two Novel Type III-Secreted Proteins of Xanthomonas campestris pv. vesicatoria Are Encoded within the hrp Pathogenicity Island

2002 ◽  
Vol 184 (5) ◽  
pp. 1340-1348 ◽  
Author(s):  
Laurent Noël ◽  
Frank Thieme ◽  
Dirk Nennstiel ◽  
Ulla Bonas
Keyword(s):  
Gene Cluster ◽  
Pseudomonas Syringae ◽  
Xanthomonas Campestris ◽  
Pathogenicity Island ◽  
Effector Proteins ◽  
In Planta ◽  
Type Iii ◽  
Reverse Transcription Pcr ◽  
New Genes ◽  
Type Iii Protein Secretion

ABSTRACT The Hrp type III protein secretion system (TTSS) is essential for pathogenicity of gram-negative plant pathogen Xanthomonas campestris pv. vesicatoria. cDNA-amplified fragment length polymorphism and reverse transcription-PCR analyses identified new genes, regulated by key hrp regulator HrpG, in the regions flanking the hrp gene cluster. Sequence analysis revealed genes encoding HpaG, a predicted leucine-rich repeat-containing protein, the lysozyme-like HpaH protein, and XopA and XopD, which are similar in sequence to Hpa1 from Xanthomonas oryzae pv. oryzae and PsvA from Pseudomonas syringae, respectively. XopA and XopD (Xanthomonas outer proteins) are secreted by the Xanthomonas Hrp TTSS and thus represent putative effector proteins. Mutations in xopA, but not in xopD, resulted in reduced bacterial growth in planta and delayed plant reactions in susceptible and resistant host plants. Since the xopD promoter contains a putative hrp box, which is characteristic of hrpL-regulated genes in P. syringae and Erwinia spp., the gene was probably acquired by horizontal gene transfer. Interestingly, the regions flanking the hrp gene cluster also contain insertion sequences and genes for a putative transposase and a tRNAArg. These features suggest that the hrp gene cluster of X. campestris pv. vesicatoria is part of a pathogenicity island.

scholarly journals Postfixation detergent treatment for immunofluorescence suppresses localization of some integral membrane proteins.

1985 ◽  
Vol 33 (8) ◽  
pp. 813-820 ◽  
Author(s):  
K L Goldenthal ◽  
K Hedman ◽  
J W Chen ◽  
J T August ◽  
M C Willingham
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Integral Membrane Protein ◽  
Membrane Glycoproteins ◽  
Animal Cells ◽  
Cytoplasmic Proteins ◽  
Membrane Antigens ◽  
Endocytic Vesicles ◽  
Triton X ◽  
Nonionic Detergents

Immunofluorescence microscopy of cultured animal cells is often performed after detergent permeabilization of formaldehyde-fixed cellular membranes so that antibodies may have access to intracellular antigens. A comparison was made of the ability of several detergents, after formaldehyde fixation, to affect localization of intracellular proteins or to permeabilize different organelles to antibodies. Saponin, a detergent-like molecule that can permeabilize cholesterol-containing membranes, was also used. Four monoclonal antibodies were found to have a bright, discrete fluorescence localization with saponin alone, but were almost undetectable when the cells were treated with nonionic detergents such as Triton X-100 or NP-40. These immunoglobulin G antibodies included two against lysosomal membrane glycoproteins, one against an integral membrane protein found in the plasma membrane and endocytic vesicles, and one against a membrane protein in the endoplasmic reticulum and the nuclear envelope. However, antigens localized in mitochondria and the nucleus required the use of a detergent such as Triton X-100 for their detection. The detection of a number of other membrane or cytoplasmic proteins was unaffected by Triton X-100 treatment. It was concluded that nonionic detergents such as Triton X-100 cause artifactual loss of detection of some membrane proteins, and saponin is a favorable alternative reagent for immunofluorescence detection of intracellular membrane antigens in many organelles.

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