TY - JOUR A1 - Gupta, Shishir K. A1 - Srivastava, Mugdha A1 - Osmanoglu, Özge A1 - Xu, Zhuofei A1 - Brakhage, Axel A. A1 - Dandekar, Thomas T1 - Aspergillus fumigatus versus genus Aspergillus: conservation, adaptive evolution and specific virulence genes JF - Microorganisms N2 - Aspergillus is an important fungal genus containing economically important species, as well as pathogenic species of animals and plants. Using eighteen fungal species of the genus Aspergillus, we conducted a comprehensive investigation of conserved genes and their evolution. This also allows us to investigate the selection pressure driving the adaptive evolution in the pathogenic species A. fumigatus. Among single-copy orthologs (SCOs) for A. fumigatus and the closely related species A. fischeri, we identified 122 versus 50 positively selected genes (PSGs), respectively. Moreover, twenty conserved genes of unknown function were established to be positively selected and thus important for adaption. A. fumigatus PSGs interacting with human host proteins show over-representation of adaptive, symbiosis-related, immunomodulatory and virulence-related pathways, such as the TGF-β pathway, insulin receptor signaling, IL1 pathway and interfering with phagosomal GTPase signaling. Additionally, among the virulence factor coding genes, secretory and membrane protein-coding genes in multi-copy gene families, 212 genes underwent positive selection and also suggest increased adaptation, such as fungal immune evasion mechanisms (aspf2), siderophore biosynthesis (sidD), fumarylalanine production (sidE), stress tolerance (atfA) and thermotolerance (sodA). These genes presumably contribute to host adaptation strategies. Genes for the biosynthesis of gliotoxin are shared among all the close relatives of A. fumigatus as an ancient defense mechanism. Positive selection plays a crucial role in the adaptive evolution of A. fumigatus. The genome-wide profile of PSGs provides valuable targets for further research on the mechanisms of immune evasion, antimycotic targeting and understanding fundamental virulence processes. KW - molecular evolution KW - phylogenetic analysis KW - adaptation KW - recombination KW - positive selection KW - human pathogenic fungi KW - genus Aspergillus KW - Aspergillus fumigatus Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-246318 SN - 2076-2607 VL - 9 IS - 10 ER - TY - JOUR A1 - Hentschel, Ute A1 - Kamke, Janine A1 - Rinke, Christian A1 - Schwientek, Patrick A1 - Mavromatis, Kostas Mavromatis A1 - Ivanova, Natalia A1 - Sczyrba, Alexander A1 - Woyke, Tanja T1 - The Candidate Phylum Poribacteria by Single-Cell Genomics: New Insights into Phylogeny, Cell-Compartmentation, Eukaryote-Like Repeat Proteins, and Other Genomic Features N2 - The candidate phylum Poribacteria is one of the most dominant and widespread members of the microbial communities residing within marine sponges. Cell compartmentalization had been postulated along with their discovery about a decade ago and their phylogenetic association to the Planctomycetes, Verrucomicrobia, Chlamydiae superphylum was proposed soon thereafter. In the present study we revised these features based on genomic data obtained from six poribacterial single cells. We propose that Poribacteria form a distinct monophyletic phylum contiguous to the PVC superphylum together with other candidate phyla. Our genomic analyses supported the possibility of cell compartmentalization in form of bacterial microcompartments. Further analyses of eukaryote-like protein domains stressed the importance of such proteins with features including tetratricopeptide repeats, leucin rich repeats as well as low density lipoproteins receptor repeats, the latter of which are reported here for the first time from a sponge symbiont. Finally, examining the most abundant protein domain family on poribacterial genomes revealed diverse phyH family proteins, some of which may be related to dissolved organic posphorus uptake. KW - Candidate Phylum Poribacteria KW - protein domains KW - genomic databases KW - phylogenetic analysis KW - genome analysis KW - sponges KW - marine bacteria KW - phylogenetics KW - polyvinyl chloride Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-112649 N1 - This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication. ER - TY - THES A1 - Tabares, Paula T1 - Antimicrobial, anti-protease and immunomodulatory activities of secondary metabolites from Caribbean sponges and their associated bacteria T1 - Sekundärmetabolite mit antimikrobiellen, Protease-hemmenden und immunmodulatorischen Aktivitäten aus karibischen Schwämmen und assoziierten Bakterien N2 - Marine sponges and their associated bacteria have been proven to be a rich source of novel secondary metabolites with therapeutic usefulness in infection and autoimmunity. This Ph.D. project aimed to isolate bioactive secondary metabolites from the marine sponges Amphimedon compressa, Aiolochroia crassa and Theonella swinhoei as well as from bacteria associated with different Caribbean sponges, specifically actinomycetes and sphingomonads. In this study, amphitoxin was isolated from the crude methanol extract of the sponge A. compressa and it was found to have antibacterial and anti-parasitic activities. Amphitoxin showed protease inhibitory activity when tested against the mammalian protease cathepsin B and the parasitic proteases rhodesain and falcipain-2. Furthermore, miraziridine A was identified in the dichloromethane extract of the sponge T. swinhoei collected offshore Israel in the Red Sea. Miraziridine A, a natural peptide isolated previously from the marine sponge Theonella aff. mirabilis, is a potent cathepsin B inhibitor with an IC50 value of 1.4 g/mL (2.1 M). Secondary metabolites from sponge-derived bacteria were also isolated and identified. A total of 79 strains belonging to 20 genera of the order Actinomycetales and seven strains belonging to two genera of the order Sphingomonadales were cultivated from 18 different Caribbean sponges and identified by 16S rRNA gene sequencing. Seven of these strains are likely to represent novel species. Crude extracts from selected strains were found to exhibit protease inhibition against cathepsins B and L, rhodesain, and falcipain-2 as well as immunomodulatory activities such as induction of cytokine release by human peripheral blood mononuclear cells. The isolates Sphingobium sp. CO105 and Lapillicoccus sp. BA53 were selected for cultivation, extraction and purification of bioactive metabolites based on initial bioactive screening results. The isoalloxazine isolumichrome was isolated from the strain Sphingobium sp. CO105 which inhibited the protease rhodesain with an IC50 of 0.2 M. The strain Lapillicoccus sp. BA53 was found to produce p-aminosalicylic acid methyl ester, which showed activity against the proteases cathepsins B and L, falcipain-2 and rhodesain. These results highlight the significance of marine sponge-associated bacteria to produce bioactive secondary metabolites with therapeutic potential in the treatment of infectious diseases and disorders of the immune system. N2 - Marine Schwämme und damit assoziierte Bakterien stellen eine wertvolle Quelle für neuartige Sekundärmetabolite mit therapeutischer Bedeutung für Infektion und Autoimmunität dar. Ziel dieser Doktorarbeit war die Isolierung bioaktiver Sekundärmetabolite aus den marinen Schwämmen Amphimedon compressa, Ailochroia crassa und Theonella swinhoei sowie von Bakterien, die mit verschiedenen karibischen Schwämmen assoziiert sind, wie z. B. Actinomyceten und Sphingomonaden. Amphotoxin wurde in dieser Studie aus dem methanolhaltigen Rohextrakt des Schwammes A. compressa isoliert. Es konnte sowohl eine antibakterielle als auch antiparasitäre Aktivität nachgewiesen werden. Der Einfluss von Amphotoxin auf die humane Protease Cathepsin B und die parasitären Proteasen Rhodesain und Falcipain-2 wurde ebenfalls getestet und es zeigte sich eine inhibitorische Wirkung gegenüber diesen Proteasen. Darüber hinaus wurde aus dem Dichlormethanextrakt des Schwammes T. swinhoei, der aus dem Roten Meer in Israel gewonnen wurde, Miraziridin A isoliert. Dieses natürliche Peptid war bereits aus dem marinen Schwamm Theonella aff. mirabilis isoliert worden. Miraziridin A ist ein starker Cathepsin B Inhibitor, der IC50 Wert beträgt 1.4 mg/mL (2.1 M). Sekundärmetabolite von aus Schwämmen gewonnenen Bakterien wurden ebenfalls isoliert und identifiziert. Es konnten 79 Stämme, die zu 20 verschiedenen Gattungen der Ordnung Actinomycetales, sowie sieben Stämme, die zu zwei Gattungen der Ordnung Sphingomonadales gehören, isoliert werden. Diese Bakterienstämme wurden aus ingesamt 18 verschiedenen karibischen Schwämmen kultiviert und mit Hilfe der 16S rRNA Sequenzierung bestimmt. Sieben dieser Stämme stellen wahrscheinlich neue Arten dar. Rohextrakte ausgewählter Stämme zeigten eine Proteasehemmung gegen die Cathepsine B und L, Rhodesain, Falcipain-2 sowie immunmodulatorische Wirkungen wie z.B. die Induktion der Cytokinfreisetzung durch menschliche periphere mononukleäre Blutzellen. Die Isolate Sphingobium sp. CO105 und Lapillicoccus sp. BA53 wurden für die Kultivierung, Extraktion und Aufreinigung von bioaktiven Metaboliten aufgrund der ersten vielversprechenden bioaktiven Testergebnisse ausgewählt. Das Isoalloxazin Isolumichrom wurde aus dem Stamm Sphingobium sp. CO105 isoliert, welches die Protease Rhodesain mit einem IC50-Wert von 0.2 M inhibiert. Für den Stamm Lapillicoccus sp. BA53 konnte nachgewiesen werden, dass er p-Aminosalicylsäuremethylester produziert, der eine Aktivität gegen die Proteasen Cathepsin B und L, Falcipain-2 und Rhodesain zeigt. Diese Ergebnisse unterstreichen die Bedeutung mariner, Schwamm-assoziierter Bakterien, die bioaktive sekundäre Metabolite mit therapeutischem Potential für die Behandlung von Infektionskrankheiten und Funktionsstörungen des Immunsystems produzieren. KW - Schwämme KW - Bakterien KW - Karibisches Meer KW - Sekundärmetabolit KW - Actinomycetes KW - sphingomonads KW - marine sponge KW - anti-protease KW - immunomodulatory KW - phylogenetic analysis Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-67000 ER - TY - JOUR A1 - Tabatabai, Julia A1 - Prifert, Christiane A1 - Pfeil, Johannes A1 - Grulich-Henn, Juergen A1 - Schnitzler, Paul T1 - Novel Respiratory Syncytial Virus (RSV) Genotype ON1 Predominates in Germany during Winter Season 2012-13 JF - PLOS ONE N2 - Respiratory syncytial virus (RSV) is the leading cause of hospitalization especially in young children with respiratory tract infections (RTI). Patterns of circulating RSV genotypes can provide a better understanding of the molecular epidemiology of RSV infection. We retrospectively analyzed the genetic diversity of RSV infection in hospitalized children with acute RTI admitted to University Hospital Heidelberg/Germany between October 2012 and April 2013. Nasopharyngeal aspirates (NPA) were routinely obtained in 240 children younger than 2 years of age who presented with clinical symptoms of upper or lower RTI. We analyzed NPAs via PCR and sequence analysis of the second variable region of the RSV G gene coding for the attachment glycoprotein. We obtained medical records reviewing routine clinical data. RSV was detected in 134/240 children. In RSV-positive patients the most common diagnosis was bronchitis/bronchiolitis (75.4%). The mean duration of hospitalization was longer in RSV-positive compared to RSV-negative patients (3.5 vs. 5.1 days; p < 0.01). RSV-A was detected in 82.1%, RSV-B in 17.9% of all samples. Phylogenetic analysis of 112 isolates revealed that the majority of RSV-A strains (65%) belonged to the novel ON1 genotype containing a 72-nucleotide duplication. However, genotype ON1 was not associated with a more severe course of illness when taking basic clinical/laboratory parameters into account. Molecular characterization of RSV confirms the co-circulation of multiple genotypes of subtype RSV-A and RSV-B. The duplication in the G gene of genotype ON1 might have an effect on the rapid spread of this emerging RSV strain. KW - group-A KW - group B genotype KW - subgroup-B KW - South Africa KW - genetic variability KW - 60-nucleotide duplication KW - molecular characterization KW - attachment glycoprotein KW - circulation patterns KW - phylogenetic analysis Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115229 VL - 9 IS - 10 ER -