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Werner E G Muller - One of the best experts on this subject based on the ideXlab platform.
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potential biological role of laccase from the sponge Suberites domuncula as an antibacterial defense component
Biochimica et Biophysica Acta, 2015Co-Authors: Xiaohong Wang, Heinz C Schroder, Michael Korzhev, Barbel Diehlseifert, Thorben Link, Muhammad Nawaz Tahir, Werner E G MullerAbstract:Abstract Background Laccases are copper-containing enzymes that catalyze the oxidation of a wide variety of phenolic substrates. Methods We describe the first poriferan laccase from the marine demosponge Suberites domuncula. Results This enzyme comprises three characteristic multicopper oxidase homologous domains. Immunohistological studies revealed that the highest expression of the laccase is in the surface zone of the animals. The expression level of the laccase gene is strongly upregulated after exposure of the animals to the bacterial endotoxin lipopolysaccharide. To allow the binding of the recombinant enzyme to ferromagnetic nanoparticles, a recombinant laccase was prepared which contained in addition to the His-tag, a Glu-tag at the N-terminus of the enzyme. The recombinant laccase was enzymatically active. The apparent Michaelis constant of the enzyme is 114 μM, using syringaldazine as substrate. Exposure of E. coli to the nanoparticles, coated with Glu-tagged laccase, and to the mediator 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) in the presence of lignin, as the oxidizable substrate, resulted in an almost complete inhibition of colony formation. Quantitative studies of the effect of the laccase-coated iron oxide nanoparticles were performed using E. coli grown in suspension in reaction tubes within a magnetic nanoparticle separator. Conclusions This newly designed magnetic nanoparticle separator allowed a removal of the nanoparticles after terminating the reaction. Using this system, a strong dose-dependent inhibition of the growth of E. coli by the laccase iron oxide nanoparticles was determined. General significance From our data we conclude that the sponge laccase is involved in the anti-bacterial defense of the sponge organism.
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Cellular Effects of Bacterial N-3-Oxo-Dodecanoyl-L-Homoserine Lactone on the Sponge Suberites domuncula (Olivi, 1792): Insights into an Intimate Inter-Kingdom Dialogue.
PLoS ONE, 2014Co-Authors: Johan Gardères, Werner E G Muller, Matthias Wiens, Joël Henry, Benoit Bernay, Andrès Ritter, Céline Zatylny-gaudin, Gaël Le PennecAbstract:Sponges and bacteria have lived together in complex consortia for 700 million years. As filter feeders, sponges prey on bacteria. Nevertheless, some bacteria are associated with sponges in symbiotic relationships. To enable this association, sponges and bacteria are likely to have developed molecular communication systems. These may include molecules such as N-acyl-L-homoserine lactones, produced by Gram-negative bacteria also within sponges. In this study, we examined the role of N-3-oxododecanoyl-L-homoserine lactone (3-oxo-C12-HSL) on the expression of immune and apoptotic genes of the host sponge Suberites domuncula. This molecule seemed to inhibit the sponge innate immune system through a decrease of the expression of genes coding for proteins sensing the bacterial membrane: a Toll-Like Receptor and a Toll-like Receptor Associated Factor 6 and for an anti-bacterial perforin-like molecule. The expression of the pro-apoptotic caspase-like 3/7 gene decreased as well, whereas the level of mRNA of anti-apoptotic genes Bcl-2 Homolog Proteins did not change. Then, we demonstrated the differential expression of proteins in presence of this 3-oxo-C12-HSL using 3D sponge cell cultures. Proteins involved in the first steps of the endocytosis process were highlighted using the 2D electrophoresis protein separation and the MALDI-TOF/TOF protein characterization: α and β subunits of the lysosomal ATPase, a cognin, cofilins-related proteins and cytoskeleton proteins actin, α tubulin and α actinin. The genetic expression of some of these proteins was subsequently followed. We propose that the 3-oxo-C12-HSL may participate in the tolerance of the sponge apoptotic and immune systems towards the presence of bacteria. Besides, the sponge may sense the 3-oxo-C12-HSL as a molecular evidence of the bacterial presence and/or density in order to regulate the populations of symbiotic bacteria in the sponge. This study is the first report of a bacterial secreted molecule acting on sponge cells and regulating the symbiotic relationship.
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hierarchical composition of the axial filament from spicules of the siliceous sponge Suberites domuncula from biosilica synthesizing nanofibrils to structure and morphology guiding triangular stems
Cell and Tissue Research, 2013Co-Authors: Werner E G Muller, Ute Schlosmacher, Heinz C Schroder, Enrico Mugnaioli, Marco Giovine, Ute Kolb, Xiaohong WangAbstract:The major structural and enzymatically active protein in spicules from siliceous sponges, e.g., for Suberites domuncula studied here, is silicatein. Silicatein has been established to be the key enzyme that catalyzes the formation of biosilica, a polymer that represents the inorganic scaffold for the spicule. In the present study, it is shown, by application of high-resolution transmission and scanning transmission electron microscopy that, during the initial phase of spicule synthesis, nanofibrils with a diameter of around 10 nm are formed that comprise bundles of between 10 and 20 nanofibrils. In intracellular vacuoles, silicasomes, the nanofibrils form polar structures with a pointed tip and a blunt end. In a time-dependent manner, these nanofibrillar bundles become embedded into a Si-rich matrix, indicative for the formation of biosilica via silicatein molecules that form the nanofibrils. These biosilicified nanofibrillar bundles become extruded from the intracellular space, where they are located in the silicasomes, to the extracellular environment by an evagination process, during which a cellular protrusion forms the axial canal in the growing spicule. The nanofibrillar bundles condense and progressively form the axial filament that becomes localized in the extracellular space. It is concluded that the silicatein-composing nanofibrils act not only as enzymatic silica bio-condensing platforms but also as a structure-giving guidance for the growing spicule.
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nocturnin in the demosponge Suberites domuncula a potential circadian clock protein controlling glycogenin synthesis in sponges
Biochemical Journal, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Xiaohong Wang, Michael Korzhev, Vlad A Grebenjuk, Ute Schlossmacher, Heinz C SchroderAbstract:Sponges are filter feeders that consume a large amount of energy to allow a controlled filtration of water through their aquiferous canal systems. It has been shown that primmorphs, three-dimensional cell aggregates prepared from the demosponge Suberites domuncula and cultured in vitro , change their morphology depending on the light supply. Upon exposure to light, primmorphs show a faster and stronger increase in DNA, protein and glycogen content compared with primmorphs that remain in the dark. The sponge genome contains nocturnin, a light/dark-controlled clock gene, the protein of which shares a high sequence similarity with the related molecule of higher metazoans. The sponge nocturnin protein was found showing a poly(A)-specific 3′-exoribonuclease activity. In addition, the cDNA of the glycogenin gene was identified for subsequent expression studies. Antibodies against nocturnin were raised and used in parallel with the cDNA to determine the regional expression of nocturnin in intact sponge specimens; the highest expression of nocturnin was seen in the epithelial layer around the aquiferous canals. Quantitative PCR analyses revealed that primmorphs after transfer from light to dark show a 10-fold increased expression in the nocturnin gene. In contrast, the expression level of glycogenin decreases in the dark by 3– 4-fold. Exposure of primmorphs to light causes a decrease in nocturnin transcripts and a concurrent increase in glycogenin transcripts. It was concluded that sponges are provided with the molecular circadian clock protein nocturnin that is highly expressed in the dark where it controls the stability of a key metabolic enzyme, glycogenin. Abbreviations: BMAL1, brain and muscle ARNT (aryl hydrocarbon receptor nuclear translocator)-like 1; CLOCK, circadian locomotor output cycles kaput; CRY, cryptochrome; Cy5, indodicarbocyanine; DAPI, 4′,6-diamidino-2-phenylindole; DIG, digoxigenin; GABA, γ-aminobutyric acid; hnRNA, heterogeneous nuclear RNA; ORF, open reading frame; Per, period; qRT–PCR, quantitative real-time reverse transcription PCR; Ror, retinoic acid-related orphan receptor; SDGAPDH, Suberites domuncula glycerol 3-phosphate dehydrogenase; SDGYG, S. domuncula glycogenin; SDNOC, S. domuncula nocturnin
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differential expression of the demosponge Suberites domuncula carotenoid oxygenases in response to light protection mechanism against the self produced toxic protein suberitine
Marine Drugs, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Michael Binder, Johannes Von Lintig, Xiaohong Wang, Heinz C SchroderAbstract:The demosponge Suberites domuncula has been described to contain high levels of a proteinaceous toxin, Suberitine, that displays haemolytic activityIn the present study this 7–8 kDa polypeptide has been isolated and was shown to exhibit also cytotoxic effects on cells of the same species. Addition of retinal, a recently identified metabolite of β-carotene that is abundantly present in S. domuncula was found to reduce both the haemolytic and the cell toxic activity of Suberitine at a molar ratio of 1:1. Spectroscopic analyses revealed that the interaction between β-carotene and Suberitine can be ascribed to a reversible energy transfer reaction. The enzyme that synthesises retinal in the sponge system is the β,β-carotene-15,15′-dioxygenase [carotene dioxygenase]. In order to clarify if this enzyme is the only β-carotene-metabolizing enzyme a further oxygenase had been identified and cloned, the (related) carotenoid oxygenase. In contrast to the dioxygenase, the carotenoid oxygenase could not degrade β-carotene or lycopene in Escherichia coli strains that produced these two carotenoids; therefore it had been termed related-carotenoid oxygenase. Exposure of primmorphs to light of different wavelengths from the visible spectrum resulted after 3 days in a strong upregulation of the dioxygenase in those 3D-cell aggregates that had been incubated with β-carotene. The strongest effect is seen with blue light at a maximum around 490 nm. It is concluded that the toxin Suberitine is non-covalently modified by retinal, the cleavage product from β-carotene via the enzyme carotene dioxygenase, a light inducible oxygenase. Hence, this study highlights that in S. domuncula the bioactive metabolite, retinal, has the property to detoxify its homologous toxin.
Heinz C Schroder - One of the best experts on this subject based on the ideXlab platform.
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potential biological role of laccase from the sponge Suberites domuncula as an antibacterial defense component
Biochimica et Biophysica Acta, 2015Co-Authors: Xiaohong Wang, Heinz C Schroder, Michael Korzhev, Barbel Diehlseifert, Thorben Link, Muhammad Nawaz Tahir, Werner E G MullerAbstract:Abstract Background Laccases are copper-containing enzymes that catalyze the oxidation of a wide variety of phenolic substrates. Methods We describe the first poriferan laccase from the marine demosponge Suberites domuncula. Results This enzyme comprises three characteristic multicopper oxidase homologous domains. Immunohistological studies revealed that the highest expression of the laccase is in the surface zone of the animals. The expression level of the laccase gene is strongly upregulated after exposure of the animals to the bacterial endotoxin lipopolysaccharide. To allow the binding of the recombinant enzyme to ferromagnetic nanoparticles, a recombinant laccase was prepared which contained in addition to the His-tag, a Glu-tag at the N-terminus of the enzyme. The recombinant laccase was enzymatically active. The apparent Michaelis constant of the enzyme is 114 μM, using syringaldazine as substrate. Exposure of E. coli to the nanoparticles, coated with Glu-tagged laccase, and to the mediator 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) in the presence of lignin, as the oxidizable substrate, resulted in an almost complete inhibition of colony formation. Quantitative studies of the effect of the laccase-coated iron oxide nanoparticles were performed using E. coli grown in suspension in reaction tubes within a magnetic nanoparticle separator. Conclusions This newly designed magnetic nanoparticle separator allowed a removal of the nanoparticles after terminating the reaction. Using this system, a strong dose-dependent inhibition of the growth of E. coli by the laccase iron oxide nanoparticles was determined. General significance From our data we conclude that the sponge laccase is involved in the anti-bacterial defense of the sponge organism.
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hierarchical composition of the axial filament from spicules of the siliceous sponge Suberites domuncula from biosilica synthesizing nanofibrils to structure and morphology guiding triangular stems
Cell and Tissue Research, 2013Co-Authors: Werner E G Muller, Ute Schlosmacher, Heinz C Schroder, Enrico Mugnaioli, Marco Giovine, Ute Kolb, Xiaohong WangAbstract:The major structural and enzymatically active protein in spicules from siliceous sponges, e.g., for Suberites domuncula studied here, is silicatein. Silicatein has been established to be the key enzyme that catalyzes the formation of biosilica, a polymer that represents the inorganic scaffold for the spicule. In the present study, it is shown, by application of high-resolution transmission and scanning transmission electron microscopy that, during the initial phase of spicule synthesis, nanofibrils with a diameter of around 10 nm are formed that comprise bundles of between 10 and 20 nanofibrils. In intracellular vacuoles, silicasomes, the nanofibrils form polar structures with a pointed tip and a blunt end. In a time-dependent manner, these nanofibrillar bundles become embedded into a Si-rich matrix, indicative for the formation of biosilica via silicatein molecules that form the nanofibrils. These biosilicified nanofibrillar bundles become extruded from the intracellular space, where they are located in the silicasomes, to the extracellular environment by an evagination process, during which a cellular protrusion forms the axial canal in the growing spicule. The nanofibrillar bundles condense and progressively form the axial filament that becomes localized in the extracellular space. It is concluded that the silicatein-composing nanofibrils act not only as enzymatic silica bio-condensing platforms but also as a structure-giving guidance for the growing spicule.
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nocturnin in the demosponge Suberites domuncula a potential circadian clock protein controlling glycogenin synthesis in sponges
Biochemical Journal, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Xiaohong Wang, Michael Korzhev, Vlad A Grebenjuk, Ute Schlossmacher, Heinz C SchroderAbstract:Sponges are filter feeders that consume a large amount of energy to allow a controlled filtration of water through their aquiferous canal systems. It has been shown that primmorphs, three-dimensional cell aggregates prepared from the demosponge Suberites domuncula and cultured in vitro , change their morphology depending on the light supply. Upon exposure to light, primmorphs show a faster and stronger increase in DNA, protein and glycogen content compared with primmorphs that remain in the dark. The sponge genome contains nocturnin, a light/dark-controlled clock gene, the protein of which shares a high sequence similarity with the related molecule of higher metazoans. The sponge nocturnin protein was found showing a poly(A)-specific 3′-exoribonuclease activity. In addition, the cDNA of the glycogenin gene was identified for subsequent expression studies. Antibodies against nocturnin were raised and used in parallel with the cDNA to determine the regional expression of nocturnin in intact sponge specimens; the highest expression of nocturnin was seen in the epithelial layer around the aquiferous canals. Quantitative PCR analyses revealed that primmorphs after transfer from light to dark show a 10-fold increased expression in the nocturnin gene. In contrast, the expression level of glycogenin decreases in the dark by 3– 4-fold. Exposure of primmorphs to light causes a decrease in nocturnin transcripts and a concurrent increase in glycogenin transcripts. It was concluded that sponges are provided with the molecular circadian clock protein nocturnin that is highly expressed in the dark where it controls the stability of a key metabolic enzyme, glycogenin. Abbreviations: BMAL1, brain and muscle ARNT (aryl hydrocarbon receptor nuclear translocator)-like 1; CLOCK, circadian locomotor output cycles kaput; CRY, cryptochrome; Cy5, indodicarbocyanine; DAPI, 4′,6-diamidino-2-phenylindole; DIG, digoxigenin; GABA, γ-aminobutyric acid; hnRNA, heterogeneous nuclear RNA; ORF, open reading frame; Per, period; qRT–PCR, quantitative real-time reverse transcription PCR; Ror, retinoic acid-related orphan receptor; SDGAPDH, Suberites domuncula glycerol 3-phosphate dehydrogenase; SDGYG, S. domuncula glycogenin; SDNOC, S. domuncula nocturnin
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differential expression of the demosponge Suberites domuncula carotenoid oxygenases in response to light protection mechanism against the self produced toxic protein suberitine
Marine Drugs, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Michael Binder, Johannes Von Lintig, Xiaohong Wang, Heinz C SchroderAbstract:The demosponge Suberites domuncula has been described to contain high levels of a proteinaceous toxin, Suberitine, that displays haemolytic activityIn the present study this 7–8 kDa polypeptide has been isolated and was shown to exhibit also cytotoxic effects on cells of the same species. Addition of retinal, a recently identified metabolite of β-carotene that is abundantly present in S. domuncula was found to reduce both the haemolytic and the cell toxic activity of Suberitine at a molar ratio of 1:1. Spectroscopic analyses revealed that the interaction between β-carotene and Suberitine can be ascribed to a reversible energy transfer reaction. The enzyme that synthesises retinal in the sponge system is the β,β-carotene-15,15′-dioxygenase [carotene dioxygenase]. In order to clarify if this enzyme is the only β-carotene-metabolizing enzyme a further oxygenase had been identified and cloned, the (related) carotenoid oxygenase. In contrast to the dioxygenase, the carotenoid oxygenase could not degrade β-carotene or lycopene in Escherichia coli strains that produced these two carotenoids; therefore it had been termed related-carotenoid oxygenase. Exposure of primmorphs to light of different wavelengths from the visible spectrum resulted after 3 days in a strong upregulation of the dioxygenase in those 3D-cell aggregates that had been incubated with β-carotene. The strongest effect is seen with blue light at a maximum around 490 nm. It is concluded that the toxin Suberitine is non-covalently modified by retinal, the cleavage product from β-carotene via the enzyme carotene dioxygenase, a light inducible oxygenase. Hence, this study highlights that in S. domuncula the bioactive metabolite, retinal, has the property to detoxify its homologous toxin.
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inducible asabf type antimicrobial peptide from the sponge Suberites domuncula microbicidal and hemolytic activity in vitro and toxic effect on molluscs in vivo
Marine Drugs, 2011Co-Authors: Matthias Wiens, Renato Batel, Heinz C Schroder, Xiaohong Wang, Michael Korzhev, Werner E G MullerAbstract:Since sponges, as typical filter-feeders, are exposed to a high load of attacking prokaryotic and eukaryotic organisms, they are armed with a wide arsenal of antimicrobial/cytostatic low-molecular-weight, non-proteinaceous bioactive compounds. Here we present the first sponge agent belonging to the group of ASABF-type antimicrobial peptides. The ASABF gene was identified and cloned from the demosponge Suberites domuncula. The mature peptide, with a length of 64 aa residues has a predicted pI of 9.24, and comprises the characteristic CSα β structural motif. Consequently, the S. domuncula ASABF shares high similarity with the nematode ASABFs; it is distantly related to the defensins. The recombinant peptide was found to display besides microbicidal activity, anti-fungal activity. In addition, the peptide lyses human erythrocytes. The expression of ASABF is upregulated after exposure to the apoptosis-inducing agent 2,2′-dipyridyl. During the process of apoptosis of surface tissue of S. domuncula, grazing gastropods (Bittium sp.) are attracted by quinolinic acid which is synthesized through the kynurenine pathway by the enzyme 3-hydroxyanthranilate 3,4-dioxygenase (HAD). Finally, the gastropods are repelled from the sponge tissue by the ASABF. It is shown that the effector peptide ASABF is sequentially expressed after the induction of the HAD gene and a caspase, as a central enzyme executing apoptosis.
Matthias Wiens - One of the best experts on this subject based on the ideXlab platform.
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Cellular Effects of Bacterial N-3-Oxo-Dodecanoyl-L-Homoserine Lactone on the Sponge Suberites domuncula (Olivi, 1792): Insights into an Intimate Inter-Kingdom Dialogue.
PLoS ONE, 2014Co-Authors: Johan Gardères, Werner E G Muller, Matthias Wiens, Joël Henry, Benoit Bernay, Andrès Ritter, Céline Zatylny-gaudin, Gaël Le PennecAbstract:Sponges and bacteria have lived together in complex consortia for 700 million years. As filter feeders, sponges prey on bacteria. Nevertheless, some bacteria are associated with sponges in symbiotic relationships. To enable this association, sponges and bacteria are likely to have developed molecular communication systems. These may include molecules such as N-acyl-L-homoserine lactones, produced by Gram-negative bacteria also within sponges. In this study, we examined the role of N-3-oxododecanoyl-L-homoserine lactone (3-oxo-C12-HSL) on the expression of immune and apoptotic genes of the host sponge Suberites domuncula. This molecule seemed to inhibit the sponge innate immune system through a decrease of the expression of genes coding for proteins sensing the bacterial membrane: a Toll-Like Receptor and a Toll-like Receptor Associated Factor 6 and for an anti-bacterial perforin-like molecule. The expression of the pro-apoptotic caspase-like 3/7 gene decreased as well, whereas the level of mRNA of anti-apoptotic genes Bcl-2 Homolog Proteins did not change. Then, we demonstrated the differential expression of proteins in presence of this 3-oxo-C12-HSL using 3D sponge cell cultures. Proteins involved in the first steps of the endocytosis process were highlighted using the 2D electrophoresis protein separation and the MALDI-TOF/TOF protein characterization: α and β subunits of the lysosomal ATPase, a cognin, cofilins-related proteins and cytoskeleton proteins actin, α tubulin and α actinin. The genetic expression of some of these proteins was subsequently followed. We propose that the 3-oxo-C12-HSL may participate in the tolerance of the sponge apoptotic and immune systems towards the presence of bacteria. Besides, the sponge may sense the 3-oxo-C12-HSL as a molecular evidence of the bacterial presence and/or density in order to regulate the populations of symbiotic bacteria in the sponge. This study is the first report of a bacterial secreted molecule acting on sponge cells and regulating the symbiotic relationship.
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nocturnin in the demosponge Suberites domuncula a potential circadian clock protein controlling glycogenin synthesis in sponges
Biochemical Journal, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Xiaohong Wang, Michael Korzhev, Vlad A Grebenjuk, Ute Schlossmacher, Heinz C SchroderAbstract:Sponges are filter feeders that consume a large amount of energy to allow a controlled filtration of water through their aquiferous canal systems. It has been shown that primmorphs, three-dimensional cell aggregates prepared from the demosponge Suberites domuncula and cultured in vitro , change their morphology depending on the light supply. Upon exposure to light, primmorphs show a faster and stronger increase in DNA, protein and glycogen content compared with primmorphs that remain in the dark. The sponge genome contains nocturnin, a light/dark-controlled clock gene, the protein of which shares a high sequence similarity with the related molecule of higher metazoans. The sponge nocturnin protein was found showing a poly(A)-specific 3′-exoribonuclease activity. In addition, the cDNA of the glycogenin gene was identified for subsequent expression studies. Antibodies against nocturnin were raised and used in parallel with the cDNA to determine the regional expression of nocturnin in intact sponge specimens; the highest expression of nocturnin was seen in the epithelial layer around the aquiferous canals. Quantitative PCR analyses revealed that primmorphs after transfer from light to dark show a 10-fold increased expression in the nocturnin gene. In contrast, the expression level of glycogenin decreases in the dark by 3– 4-fold. Exposure of primmorphs to light causes a decrease in nocturnin transcripts and a concurrent increase in glycogenin transcripts. It was concluded that sponges are provided with the molecular circadian clock protein nocturnin that is highly expressed in the dark where it controls the stability of a key metabolic enzyme, glycogenin. Abbreviations: BMAL1, brain and muscle ARNT (aryl hydrocarbon receptor nuclear translocator)-like 1; CLOCK, circadian locomotor output cycles kaput; CRY, cryptochrome; Cy5, indodicarbocyanine; DAPI, 4′,6-diamidino-2-phenylindole; DIG, digoxigenin; GABA, γ-aminobutyric acid; hnRNA, heterogeneous nuclear RNA; ORF, open reading frame; Per, period; qRT–PCR, quantitative real-time reverse transcription PCR; Ror, retinoic acid-related orphan receptor; SDGAPDH, Suberites domuncula glycerol 3-phosphate dehydrogenase; SDGYG, S. domuncula glycogenin; SDNOC, S. domuncula nocturnin
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differential expression of the demosponge Suberites domuncula carotenoid oxygenases in response to light protection mechanism against the self produced toxic protein suberitine
Marine Drugs, 2012Co-Authors: Werner E G Muller, Matthias Wiens, Michael Binder, Johannes Von Lintig, Xiaohong Wang, Heinz C SchroderAbstract:The demosponge Suberites domuncula has been described to contain high levels of a proteinaceous toxin, Suberitine, that displays haemolytic activityIn the present study this 7–8 kDa polypeptide has been isolated and was shown to exhibit also cytotoxic effects on cells of the same species. Addition of retinal, a recently identified metabolite of β-carotene that is abundantly present in S. domuncula was found to reduce both the haemolytic and the cell toxic activity of Suberitine at a molar ratio of 1:1. Spectroscopic analyses revealed that the interaction between β-carotene and Suberitine can be ascribed to a reversible energy transfer reaction. The enzyme that synthesises retinal in the sponge system is the β,β-carotene-15,15′-dioxygenase [carotene dioxygenase]. In order to clarify if this enzyme is the only β-carotene-metabolizing enzyme a further oxygenase had been identified and cloned, the (related) carotenoid oxygenase. In contrast to the dioxygenase, the carotenoid oxygenase could not degrade β-carotene or lycopene in Escherichia coli strains that produced these two carotenoids; therefore it had been termed related-carotenoid oxygenase. Exposure of primmorphs to light of different wavelengths from the visible spectrum resulted after 3 days in a strong upregulation of the dioxygenase in those 3D-cell aggregates that had been incubated with β-carotene. The strongest effect is seen with blue light at a maximum around 490 nm. It is concluded that the toxin Suberitine is non-covalently modified by retinal, the cleavage product from β-carotene via the enzyme carotene dioxygenase, a light inducible oxygenase. Hence, this study highlights that in S. domuncula the bioactive metabolite, retinal, has the property to detoxify its homologous toxin.
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inducible asabf type antimicrobial peptide from the sponge Suberites domuncula microbicidal and hemolytic activity in vitro and toxic effect on molluscs in vivo
Marine Drugs, 2011Co-Authors: Matthias Wiens, Renato Batel, Heinz C Schroder, Xiaohong Wang, Michael Korzhev, Werner E G MullerAbstract:Since sponges, as typical filter-feeders, are exposed to a high load of attacking prokaryotic and eukaryotic organisms, they are armed with a wide arsenal of antimicrobial/cytostatic low-molecular-weight, non-proteinaceous bioactive compounds. Here we present the first sponge agent belonging to the group of ASABF-type antimicrobial peptides. The ASABF gene was identified and cloned from the demosponge Suberites domuncula. The mature peptide, with a length of 64 aa residues has a predicted pI of 9.24, and comprises the characteristic CSα β structural motif. Consequently, the S. domuncula ASABF shares high similarity with the nematode ASABFs; it is distantly related to the defensins. The recombinant peptide was found to display besides microbicidal activity, anti-fungal activity. In addition, the peptide lyses human erythrocytes. The expression of ASABF is upregulated after exposure to the apoptosis-inducing agent 2,2′-dipyridyl. During the process of apoptosis of surface tissue of S. domuncula, grazing gastropods (Bittium sp.) are attracted by quinolinic acid which is synthesized through the kynurenine pathway by the enzyme 3-hydroxyanthranilate 3,4-dioxygenase (HAD). Finally, the gastropods are repelled from the sponge tissue by the ASABF. It is shown that the effector peptide ASABF is sequentially expressed after the induction of the HAD gene and a caspase, as a central enzyme executing apoptosis.
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Interaction of the retinoic acid signaling pathway with spicule formation in the marine sponge Suberites domuncula through activation of bone morphogenetic protein-1
Biochimica et biophysica acta, 2011Co-Authors: Werner E G Muller, Matthias Wiens, Michael Binder, Johannes Von Lintig, Yue-wei Guo, Xiaohong Wang, Jaap A. Kaandorp, Heinz C SchroderAbstract:Abstract Background The formation of the spicules in siliceous sponges involves the formation of cylinder-like structures in the extraspicular space, composed of the enzyme silicatein and the calcium-dependent lectin. Scope of review Molecular cloning of the cDNAs (carotene dioxygenase, retinal dehydrogenase, and BMB-1 [bone morphogenic protein-1]) from the demosponge Suberites domuncula was performed. These tools were used to understand the retinoid metabolism in the animal by qRT-PCR, immunoblotting and TEM. Major conclusions We demonstrate that silintaphin-2, a silicatein-interacting protein, is processed from a longer-sized 15-kDa precursor to a truncated, shorter-sized 13 kDa calcium-binding protein via proteolytic cleavage at the dipeptide Ala↓Asp, mediated by BMP-1. The expression of this protease as well as the expression of two key enzymes of the carotinoid metabolism, the β,β-carotene-15,15′-dioxygenase and the retinal dehydrogenase/reductase, were found to be strongly up-regulated by retinoic acid. Hence retinoic acid turned out to be a key factor in skeletogenesis in the most ancient still existing metazoans, the sponges. General significance It is shown that retinoic acid regulates the formation of the organic cylinder that surrounds the axis of the spicules and enables, as a scaffold, the radial apposition of new silica layers and hence the growth of the spicules.
Renato Batel - One of the best experts on this subject based on the ideXlab platform.
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inducible asabf type antimicrobial peptide from the sponge Suberites domuncula microbicidal and hemolytic activity in vitro and toxic effect on molluscs in vivo
Marine Drugs, 2011Co-Authors: Matthias Wiens, Renato Batel, Heinz C Schroder, Xiaohong Wang, Michael Korzhev, Werner E G MullerAbstract:Since sponges, as typical filter-feeders, are exposed to a high load of attacking prokaryotic and eukaryotic organisms, they are armed with a wide arsenal of antimicrobial/cytostatic low-molecular-weight, non-proteinaceous bioactive compounds. Here we present the first sponge agent belonging to the group of ASABF-type antimicrobial peptides. The ASABF gene was identified and cloned from the demosponge Suberites domuncula. The mature peptide, with a length of 64 aa residues has a predicted pI of 9.24, and comprises the characteristic CSα β structural motif. Consequently, the S. domuncula ASABF shares high similarity with the nematode ASABFs; it is distantly related to the defensins. The recombinant peptide was found to display besides microbicidal activity, anti-fungal activity. In addition, the peptide lyses human erythrocytes. The expression of ASABF is upregulated after exposure to the apoptosis-inducing agent 2,2′-dipyridyl. During the process of apoptosis of surface tissue of S. domuncula, grazing gastropods (Bittium sp.) are attracted by quinolinic acid which is synthesized through the kynurenine pathway by the enzyme 3-hydroxyanthranilate 3,4-dioxygenase (HAD). Finally, the gastropods are repelled from the sponge tissue by the ASABF. It is shown that the effector peptide ASABF is sequentially expressed after the induction of the HAD gene and a caspase, as a central enzyme executing apoptosis.
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tubulin polymerization promoting protein tppp ortholog from Suberites domuncula and comparative analysis of tppp p25 gene family
Biologia, 2011Co-Authors: Mauro Stifanic, Renato Batel, Werner E G MullerAbstract:Sponges are one of the oldest metazoan phyla that are, due to their highly conservative nature, often referred to as the living fossils of multicellular animals. As such, they are a very important model for evolutionary, developmental and functional studies of Metazoa. Tubulin polymerization promoting proteins (TPPPs) are defined by the presence of p25-alpha domain (Pfam05517). Their functional characteristics resemble those of microtubule-associated proteins. Presence of TPPP homologous genes has been postulated in all eukaryotes with ciliated cells and their primary function has been proposed as some basic cilia-connected function. We present here the genomic structure and the corresponding cDNA sequence of one poriferan TPPP homolog (SdTPPP) isolated from the marine sponge Suberites domuncula; and a comparative analysis of TPPP homolog sequences and genomic structures from other Eukaryotes. Our results confirm the radiation of one TPPP homolog into three distinct genes in the Vertebrate lineage, but the origin of different sequences and their phylogenetic relationships show to be influenced by alternative protein isoforms, independent gene duplications, modularity of the p25-alpha domain and possible adaptational requirements to environmental conditions.
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isolation and characterization of a mn ii oxidizing bacillus strain from the demosponge Suberites domuncula
Marine Drugs, 2010Co-Authors: Xiaohong Wang, Renato Batel, Heinz C Schroder, Matthias Wiens, Vladislav A Grebenjuk, Mugdha Divekar, Werner E G MullerAbstract:In this study we demonstrate that the demosponge Suberites domuncula harbors a Mn(II)-oxidizing bacterium, a Bacillus strain, termed BAC-SubDo-03. Our studies showed that Mn(II) stimulates bacterial growth and induces sporulation. Moreover, we show that these bacteria immobilize manganese on their cell surface. Comparison of the 16S rDNA sequence allowed the grouping of BAC-SubDo-03 to the Mn-precipitating bacteria. Analysis of the spore cell wall revealed that it contains an Mn(II)-oxidizing enzyme. Co-incubation studies of BAC-SubDo-03 with 100 μM MnCl2 and >1 μM of CuCl2 showed an increase in their Mn(II)-oxidizing capacity. In order to prove that a multicopper oxidase-like enzyme(s) (MCO) exists in the cell wall of the S. domuncula-associated BAC-SubDo-03 Bacillus strain, the gene encoding this enzyme was cloned (mnxG-SubDo-03). Sequence alignment of the deduced MCO protein (MnxG-SubDo-03) revealed that the sponge bacterium clusters together with known Mn(II)-oxidizing bacteria. The expression of the mnxG-SubDo-03 gene is under strong control of extracellular Mn(II). Based on these findings, we assume that BAC-SubDo-03 might serve as a Mn reserve in the sponge providing the animal with the capacity to detoxify Mn in the environment. Applying the in vitro primmorph cell culture system we could demonstrate that sponge cells, that were co-incubated with BAC-SubDo-03 in the presence of Mn(II), show an increased proliferation potential.
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contribution to the ecology of the sponge Suberites domuncula olivi 1792 a field study northen adriatic sea croatia
Periodicum Biologorum, 2007Co-Authors: Andrej Jaklin, Franz Brümmer, Bojan Hamer, Rudolf K Zahn, Dijana Pavicichamer, Martin Pfannkuchen, E Werner G Muller, Renato BatelAbstract:Background and purpose: Through centuries sponges and other marine invertebrates have been widely affected by their use in commercial trade and for human benefit, including usage for public aquaria and in scientific work. Sponge populations are also impacted by bottom fishing and by pollution. Due to the limited knowledge about the extent of these factors the main objective of this study was to estimate the population abundance and colour variations of the sponge species Suberites domuncula near Rovinj (northern Adriatic Sea, Croatia). Materials and methods: Specimens of S. domuncula were studied in situ and collected by SCUBA diving at a site about 2.7 km off Rovinj along a 3 x 50 m transect (32.8-33.5 m depth). Results and conclusions: During a 16 month period from June 2004 to September 2005, 768 specimens in total have been recorded. We determined for the first time abundance of S. domuncula as one individual per 2.8± ; ; 0.9 m2. During the research period we observed sponges of different colours: red-orange, blue, yellow, brown, and white as well as combinations thereof with differing patterns. There is a great prevalence of the red-orange colour variety of 85.2± ; ; 4.4%. S. domuncula, with an abundance of 34 to 96 specimens for 150 m2 at investigated site does not appear as a threatened species, moreover, according to the data given by Zavodnik and Simunovic (28), it appears more common in Rovinj area than in the rest of the northern Adriatic.
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Contribution to the ecology of the sponge Suberites domuncula (Olivi, 1792) (Porifera, Demospongiae, Hadromerida): Sponge contraction
Fresenius Environmental Bulletin, 2007Co-Authors: Bojan Hamer, Franz Brümmer, Andrej Jaklin, Dijana Pavičić-hamer, E.g. Werner Müller, K. Rudolf Zahn, Renato BatelAbstract:Autecology of most sponge species, including Suberites domuncula in the Adriatic Sea, is still unknown, as there are gaps in our knowledge of, for example, growth, migration, behavior, and symbiosis. The main objective of this study was to compile additional contributions on the contraction phenomena of S. domuncula. Specimens of S. domuncula were studied in situ and collected by a SCUBA diver at a site 2.7 km off Rovinj, northern Adriatic Sea, Croatia. We determined and compared, for the first time, the dimensions of sponge specimens in situ (>30 m depth) and in aquaria. Within 1hr after collection, contractions resulted in a decrease of volumes between 40.7 to 69.5%, with an average contraction of 52.1 ± 12.0%. Ex situ, permanent contractions to approximately half their size can change the consistency of the sponge, form, canal structure, water flow, including the disappearance of oscula. It is obvious, that this adaptive, stress protective phenomenon can have far-reaching consequences on the maintenance and life of S. domuncula specimens in aquaria. In the wild, in situ reversible contractions could have protective function by reducing water filtration, decreasing the input of harmful substances, and providing a mechanical strength shield against predators.
Isabel M. Müller - One of the best experts on this subject based on the ideXlab platform.
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cloning and expression of a tauropine dehydrogenase from the marine sponge Suberites domuncula
Marine Biology, 2008Co-Authors: Bruna Plese, Isabel M. Müller, Heinz C Schroder, Hans J Breter, Vladislav A Grebenjuk, Werner E G MullerAbstract:The cDNA sequence coding for tauropine dehydrogenase (TaDH) [belonging to the family of opine dehydrogenases] has been determined. Using the demosponge Suberites domuncula, we describe for the first time the tauropine dehydrogenase gene (of length 2,992 kb) from a eukaryote, consisting of two introns flanked by three exons. Moreover, two allelic variants have been identified, which are present in the different specimens either in a homozygotic or in a heterozygotic way; the data suggest an intermediary type of heritance. Phylogenetic analyses indicate that S. domuncula TaDH is only distantly related to the opine dehydrogenases from marine invertebrates; rather it comprises high sequence similarity to bacterial ornithine cyclodeaminases (OCD). In addition, expression studies revealed that the steady-state level of TaDH dropped drastically in animals, which had been exposed to elevated aeration. Antibodies raised against the recombinant sponge TaDH were used to demonstrate that S. domuncula expresses high levels of this enzyme in almost all cells. If tissue samples were kept under additional aeration no immuno-signals could be identified. A strong accumulation of the enzyme was seen around the bacteria, existing in bacteriocytes, indicating that under aerobic conditions the bacteria might produce taurine. These data suggest involvement of the sponge TaDH in the final step of the glycolytic pathway, more specifically, in regeneration of NAD(+) under anaerobic conditions. Furthermore, potential mutual influences between bacteria and sponge are discussed, claiming a horizontal gene transfer of the gene from a bacterium to the sponge.
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co expression and functional interaction of silicatein with galectin matrix guided formation of siliceous spicules in the marine demosponge Suberites domuncula
Journal of Biological Chemistry, 2006Co-Authors: Heinz C Schroder, Alexandra Boreiko, Isabel M. Müller, Wolfgang Tremel, Michael Korzhev, Hiroshi Ushijima, Carsten Eckert, Muhammad Nawaz Tahir, Werner E G MullerAbstract:Abstract Sponges (phylum Porifera) of the class of Demospongiae are stabilized by a siliceous skeleton. It is composed of silica needles (spicules), which provide the morphogenetic scaffold of these metazoans. In the center of the spicules there is an axial filament that consists predominantly of silicatein, an enzyme that catalyzes the synthesis of biosilica. By differential display of transcripts we identified additional proteins involved in silica formation. Two genes were isolated from the marine demosponge Suberites domuncula; one codes for a galectin and the other for a fibrillar collagen. The galectin forms aggregates to which silicatein molecules bind. The extent of the silicatein-mediated silica formation strongly increased if associated with the galectin. By applying a new and mild extraction procedure that avoids hydrogen fluoride treatment, native axial filaments were extracted from spicules of S. domuncula. These filaments contained, in addition to silicatein, the galectin and a few other proteins. Immunogold electron microscopic studies underscored the role of these additional proteins, in particular that of galectin, in spiculogenesis. Galectin, in addition to silicatein, presumably forms in the axial canal as well as on the surface of the spicules an organized net-like matrix. In the extraspicular space most of these complexes are arranged concentrically around the spicules. Taken together, these additional proteins, working together with silicatein, may also be relevant for potential (nano)-biotechnological applications of silicatein in the formation of surface coatings. Finally, we propose a scheme that outlines the matrix (galectin/silicatein)-guided appositional growth of spicules through centripetal and centrifugal synthesis and deposition of biosilica.
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selenium affects biosilica formation in the demosponge Suberites domuncula
FEBS Journal, 2005Co-Authors: Werner E G Muller, Anatoli Krasko, Isabel M. Müller, David Brandt, Bojan Hamer, Alexandra Borejko, Ronald Osinga, Hiroshi Ushijima, Cao Xupeng, Heinz C SchroderAbstract:Selenium is a trace element found in freshwater and the marine environment. We show that it plays a major role in spicule formation in the demosponge Suberites domuncula. If added to primmorphs, an in vitro sponge cell culture system, it stimulates the formation of siliceous spicules. Using differential display of transcripts, we demonstrate that, after a 72-h exposure of primmorphs to selenium, two genes are up-regulated; one codes for selenoprotein M and the other for a novel spicule-associated protein. The deduced protein sequence of selenoprotein M (14 kDa) shows characteristic features of metazoan selenoproteins. The spicule-associated protein (26 kDa) comprises six characteristic repeats of 20 amino acids, composed of 10 distinct hydrophobic regions (? 9 amino acids in length). Recombinant proteins were prepared, and antibodies were raised against these two proteins. Both were found to stain the central axial filament, which compri-ses the silicatein, as well as the surface of the spicules. In the presence of selenium, only the genes for selenoprotein M and spicule-associated protein are up-regulated, whereas the expression of the silicatein gene remains unchanged. Finally we show that, in the presence of selenium, larger silica aggregates are formed. We conclude that selenium has a stimulatory effect on the formation of siliceous spicules in sponges, and it may be involved in the enzymatic synthesis of biosilica components.
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innate immune defense of the sponge Suberites domuncula against bacteria involves a myd88 dependent signaling pathway induction of a perforin like molecule
Journal of Biological Chemistry, 2005Co-Authors: Matthias Wiens, Anatoli Krasko, Isabel M. Müller, Narsinh L Thakur, Hans J Breter, Michael Korzhev, Sanja Perovicottstadt, Barbel Diehlseifert, Hiroshi Ushijima, Werner E G MullerAbstract:Sponges (phylum Porifera) are the phylogenetically oldest metazoa; as filter feeders, they are abundantly exposed to marine microorganisms. Here we present data indicating that the demosponge Suberites domuncula is provided with a recognition system for Gram-negative bacteria. The lipopolysaccharide (LPS)-interacting protein was identified as a receptor on the sponge cell surface, which recognizes the bacterial endotoxin LPS. The cDNA was isolated, and the protein (Mr 49,937) was expressed. During binding to LPS, the protein dimerizes and interacts with MyD88, which was also identified and cloned. The sponge MyD88 (Mr 28,441) is composed of two protein interaction domains, a Toll/interleukin-1 receptor domain (found in MyD88 and in Toll-like receptors) and a death domain (present in MyD88 and interleukin-1 receptor-associated kinase). Northern blot experiments and in situ hybridization studies showed that after LPS treatment, the level of the LPS-interacting protein remains unchanged, whereas MyD88 is strongly up-regulated. A perforin-like molecule (Mr 74,171), the macrophage-expressed protein, was identified as an executing molecule of this pathway. This gene is highly expressed after LPS treatment, especially at the surfaces of the animals. The recombinant protein possesses biological activity and eliminates Gram-negative bacteria; it is inactive against Gram-positive bacteria. These data indicate that S. domuncula is provided with an innate immune system against Gram-negative bacteria; the ligand LPS (a pathogen-associated molecular pattern) is recognized by the pattern recognition receptor (LPS-interacting protein), which interacts with MyD88. A signal transduction is established, which results in an elevated expression of MyD88 as well as of the macrophage-expressed protein as an executing protein.
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biosilica formation in spicules of the sponge Suberites domuncula synchronous expression of a gene cluster
Genomics, 2005Co-Authors: Heinz C Schroder, Isabel M. Müller, Vladislav A Grebenjuk, Sanja Perovicottstadt, Sylvia Engel, Werner E G MullerAbstract:The formation of spicules is a complicated morphogenetic process in sponges (phylum Porifera). The primmorph system was used to demonstrate that in the demosponge Suberites domuncula the synthesis of the siliceous spicules starts intracellularly and is dependent on the concentration of silicic acid. To understand spicule formation, a cluster of genes was isolated. In the center of this cluster is the silicatein gene, which codes for the enzyme that synthesizes spicules. This gene is flanked by an ankyrin repeat gene at one side and by a tumor necrosis factor receptor-associated factor and a protein kinase gene at the other side. All genes are strongly expressed in primmorphs and intact animals after exposure to silicic acid, and this expression is restricted to those areas where the spicule formation starts or where spicules are maintained in the animals. Our observations suggest that in S. domuncula a coordinated expression of physically linked genes is essential for the synthesis of the major skeletal elements.
