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Metabolic differences between symbiont subpopulations in the deep-sea tubeworm Riftia pachyptila
(2020)
Hinzke, Tjorven ; Kleiner, Manuel ; Meister, Mareike ; Schlüter, Rabea ; Hentschker, Christian ; Pané-Farré, Jan ; Hildebrandt, Petra ; Felbeck, Horst ; Sievert, Stefan M. ; Bonn, Florian ; Völker, Uwe ; Becher, Dörte ; Schweder, Thomas ; Markert, Stephanie
The hydrothermal vent tube worm Riftia pachyptila lives in intimate symbiosis with intracellular sulfur-oxidizing gammaproteobacteria. Although the symbiont population consists of a single 16S rRNA phylotype, bacteria in the same host animal exhibit a remarkable degree of metabolic diversity: They simultaneously utilize two carbon fixation pathways and various energy sources and electron acceptors. Whether these multiple metabolic routes are employed in the same symbiont cells, or rather in distinct symbiont subpopulations, was unclear. As Riftia symbionts vary considerably in cell size and shape, we enriched individual symbiont cell sizes by density gradient centrifugation in order to test whether symbiont cells of different sizes show different metabolic profiles. Metaproteomic analysis and statistical evaluation using clustering and random forests, supported by microscopy and flow cytometry, strongly suggest that Riftia symbiont cells of different sizes represent metabolically dissimilar stages of a physiological differentiation process: Small symbionts actively divide and may establish cellular symbiont-host interaction, as indicated by highest abundance of the cell division key protein FtsZ and highly abundant chaperones and porins in this initial phase. Large symbionts, on the other hand, apparently do not divide, but still replicate DNA, leading to DNA endoreduplication. Highest abundance of enzymes for CO2 fixation, carbon storage and biosynthesis in large symbionts indicates that in this late differentiation stage the symbiont’s metabolism is efficiently geared towards the production of organic material. We propose that this division of labor between smaller and larger symbionts benefits the productivity of the symbiosis as a whole.
Dietrich, Helge Markus ; Righetto, Ricardo Diogo ; Kumar, Anuj ; Wietrzynski, Wojciech ; Trischler, Raphael ; Schuller, Sandra K. ; Wagner, Jonathan ; Schwarz, Fabian M. ; Engel, Benjamin D. ; Müller, Volker ; Schuller, Jan Michael
Filamentous enzymes have been found in all domains of life, but the advantage of filamentation is often elusive1. Some anaerobic, autotrophic bacteria have an unusual filamentous enzyme for CO2 fixation—hydrogen-dependent CO2 reductase (HDCR)2,3—which directly converts H2 and CO2 into formic acid. HDCR reduces CO2 with a higher activity than any other known biological or chemical catalyst4,5, and it has therefore gained considerable interest in two areas of global relevance: hydrogen storage and combating climate change by capturing atmospheric CO2. However, the mechanistic basis of the high catalytic turnover rate of HDCR has remained unknown. Here we use cryo-electron microscopy to reveal the structure of a short HDCR filament from the acetogenic bacterium Thermoanaerobacter kivui. The minimum repeating unit is a hexamer that consists of a formate dehydrogenase (FdhF) and two hydrogenases (HydA2) bound around a central core of hydrogenase Fe-S subunits, one HycB3 and two HycB4. These small bacterial polyferredoxin-like proteins oligomerize through their C-terminal helices to form the backbone of the filament. By combining structure-directed mutagenesis with enzymatic analysis, we show that filamentation and rapid electron transfer through the filament enhance the activity of HDCR. To investigate the structure of HDCR in situ, we imaged T. kivui cells with cryo-electron tomography and found that HDCR filaments bundle into large ring-shaped superstructures attached to the plasma membrane. This supramolecular organization may further enhance the stability and connectivity of HDCR to form a specialized metabolic subcompartment within the cell.
Hinzke, Tjorven ; Kleiner, Manuel ; Meister, Mareike ; Schlüter, Rabea ; Hentschker, Christian ; Pané-Farré, Jan ; Hildebrandt, Petra ; Felbeck, Horst ; Sievert, Stefan M. ; Bonn, Florian ; Völker, Uwe ; Becher, Dörte ; Schweder, Thomas ; Markert, Stephanie
The hydrothermal vent tubeworm Riftia pachyptila hosts a single 16S rRNA phylotype of intracellular sulfur-oxidizing symbionts, which vary considerably in cell morphology and exhibit a remarkable degree of physiological diversity and redundancy, even in the same host. To elucidate whether multiple metabolic routes are employed in the same cells or rather in distinct symbiont subpopulations, we enriched symbionts according to cell size by density gradient centrifugation. Metaproteomic analysis, microscopy, and flow cytometry strongly suggest that Riftia symbiont cells of different sizes represent metabolically dissimilar stages of a physiological differentiation process: While small symbionts actively divide and may establish cellular symbiont-host interaction, large symbionts apparently do not divide, but still replicate DNA, leading to DNA endoreduplication. Moreover, in large symbionts, carbon fixation and biomass production seem to be metabolic priorities. We propose that this division of labor between smaller and larger symbionts benefits the productivity of the symbiosis as a whole.
Reifarth, René ; Altstadt, Sebastian ; Göbel, Kathrin ; Heftrich, Tanja ; Heil, Michael ; Koloczek, Alexander ; Langer, Christoph ; Plag, Ralf ; Pohl, M. ; Sonnabend, Kerstin ; Weigand, Mario ; Adachi, T. ; Aksouh, Farouk ; Al-Khalili, J. ; AlGarawi, M. ; AlGhamdi, S. ; Alkhazov, G. ; Alkhomashi, N. ; Alvarez-Pol, Héctor ; Alvarez-Rodriguez, R. ; Andreev, V. ; Andrei, B. ; Atar, Leyla ; Aumann, Thomas ; Avdeichikov, Vladimir ; Bacri, C. ; Bagchi, S. ; Barbieri, Carlo ; Beceiro, Saul ; Beck, C. ; Beinrucker, Clemens ; Belier, Gilbert ; Bemmerer, Daniel ; Bendel, M. ; Benlliure Anaya, Jose Fernando ; Benzoni, G. ; Berjillos, R. ; Bertini, D. ; Bertulani, Carlos A. ; Bishop, S. ; Blasi, N. ; Bloch, T. ; Blumenfeld, Y. ; Bonaccorso, A. ; Boretzky, Konstanze ; Botvina, Alexander ; Boudard, A. ; Boutachkov, P. ; Boztosun, I. ; Bracco, A. ; Brambilla, S. ; Monago, J.Briz ; Caamano, Manuel ; Caesar, Christoph ; Camera, F. ; Casarejos Ruiz, Enrique ; Catford, Wilton N. ; Cederkäll, Joakim ; Cederwall, B. ; Chartier, Marielle ; Chatillon, Audrey ; Cherciu, M. ; Chulkov, Leonid V. ; Coleman-Smith, P. ; Cortina Gil, Dolores ; Crespi, F. ; Crespo, Raquel ; Cresswell, J. ; Csatlós, Margit ; Déchery, F. ; Davids, B. ; Davinson, Thomas ; Derya, Vera ; Detistov, P. ; Díaz Fernández, Paloma ; DiJulio, D. ; Dmitry, S. ; Doré, D. ; Dueñas, J. ; Dupont, Emmeric ; Egelhof, Peter ; Egorova, I. ; Elekes, Zoltán ; Enders, Joachim ; Endres, Janis ; Ershov, S. ; Ershova, Olga ; Fernández Domínguez, Beatriz ; Fetisov, A. ; Fiori, Enrico ; Fomichev, A. ; Fonseca, M. ; Fraile, L. ; Freer, Martin ; Friese, Jürgen ; Garcia Borge, Maria José ; Galaviz Redondo, Daniel ; Gannon, S. ; Garg, U. ; Gašparić, Igor ; Gasques, L. ; Gastineau, B. ; Geissel, Hans ; Gernhäuser, Roman ; Ghosh, T. ; Gilbert, M. ; Glorius, Jan ; Golubev, Pavel ; Gorshkov, Alexander ; Gourishetty, A. ; Grigorenko, Leonid V. ; Gulyas, J. ; Haiduc, Maria ; Hammache, F. ; Harakeh, Muhsin N. ; Hass, M. ; Heine, Marcel ; Hennig, Andreas ; Henriques, Ana ; Herzberg, Rolf-Dietmar ; Holl, Matthias ; Ignatov, Alexander ; Ignatyuk, A. ; Ilieva, Stoyanka ; Ivanov, Marian ; Iwasa, N. ; Jakobsson, B. ; Johansson, Håkan T. ; Jonson, Björn ; Joshi, P. ; Junghans, Arnd Rudolf ; Jurado Apruzzese, Beatriz ; Körner, G. ; Kalantar, N. ; Kanungo, Rituparna ; Kelic-Heil, Aleksandra ; Kezzar, K. ; Khan, E. ; Khanzadeev, Alexei ; Kiselev, Oleg A. ; Kogimtzis, M. ; Körper, D. ; Kräckmann, S. ; Kröll, Thorsten ; Krücken, Reiner ; Krasznahorkay, Attila ; Kratz, Jens-Volker ; Kresan, Dmytro ; Krings, Thomas ; Krumbholz, A. ; Krupko, S. ; Kulessa, Reinhard ; Kumar, Shyam ; Kurz, Nikolaus ; Kuzmin, E. ; Labiche, Marc ; Langanke, Karlheinz ; Lazarus, I. ; Bleis, T.Le ; Lederer, Claudia ; Lemasson, A. ; Lemmon, Roy Crawford ; Liberati, V. ; Litvinov, Yuri A. ; Löher, Bastian ; Herraiz, J.Lopez ; Münzenberg, G. ; Machado, Jorge ; Maev, E. ; Mahata, Kripamay ; Mancusi, D. ; Marganiec, Justyna ; Perez, M.Martinez ; Marusov, V. ; Mengoni, D. ; Million, B. ; Morcelle, V. ; Moreno, O. ; Movsesyan, Alina ; Nácher González, Enrique ; Najafi, Mohammad Ali ; Nakamura, T. ; Naqvi, F. ; Nikolski, E. ; Nilsson, Thomas ; Nociforo, Chiara ; Nolan, P. ; Novatsky, B. ; Nyman, Göran Hugo ; Ornelas, A. ; Palit, R. ; Pandit, S. ; Panin, Valerii ; Paradela Dobarro, Carlos ; Parkar, V. ; Paschalis, Stefanos ; Pawłowski, P. ; Perea Martinez, Angel ; Pereira, J. ; Petrache, C. ; Petri, Marina ; Pickstone, Simon Glynn ; Pietralla, Norbert ; Pietri, Stephane ; Pivovarov, Y. ; Potlog, P. ; Prokofiev, Andrey ; Rastrepina, G. ; Rauscher, Thomas ; Ribeiro Jimenez, Guillermo ; Ricciardi, Maria Valentina ; Richter, Achim ; Rigollet, Catherine ; Riisager, Karsten ; Rios, A. ; Ritter, C. ; Rodríguez Frutos, Tomás Raúl ; Rodríguez Vignote, Javier ; Röder, Marko ; Romig, Christopher ; Rossi, Dominic M. ; Roussel-Chomaz, P. ; Rout, Prakash Chandra ; Roy, S. ; Söderström, P. ; Saha Sarkar, Maitreyee ; Sakuta, S. ; Salsac, M. ; Sampson, J. ; Sanchez, J. ; Saez, del Rio ; Rosado, J.Sanchez ; Sanjari, Mohammad Shahab ; Sarriguren, P. ; Sauerwein, Anne ; Savran, Deniz ; Scheidenberger, Christoph ; Scheit, Heiko ; Schmidt, Stefan ; Schmitt, C. ; Schnorrenberger, Linda ; Schrock, P. ; Schwengner, R. ; Seddon, D. ; Sherrill, B. ; Shrivastava, A. ; Sidorchuk, S. ; Silva, Joel ; Simon, Haik ; Simpson, E. ; Singh, P. ; Slobodan, D. ; Sohler, D. ; Spieker, Mark-Christoph ; Stach, D. ; Stan, E. ; Stanoiu, M. ; Stepantsov, S. ; Stevenson, P. ; Strieder, F. ; Stuhl, László ; Suda, T. ; Sümmerer, Klaus ; Streicher, Branislav ; Taieb, Julien ; Takechi, M. ; Tanihata, I. ; Taylor, J. ; Tengblad, Olof ; Ter-Akopian, G. ; Terashima, S. ; Teubig, P. ; Thies, Ronja ; Thoennessen, M. ; Thomas, T. ; Thornhill, J. ; Thungstrom, G. ; Timar, J. ; Togano, Yasuhiro ; Tomohiro, U. ; Tornyi, T. ; Tostevin, J. ; Townsley, C. ; Trautmann, Wolfgang ; Trivedi, T. ; Typel, Stefan ; Uberseder, E. ; Udias, J. ; Uesaka, T. ; Uvarov, L. ; Vajta, Z. ; Velho, Paulo ; Vikhrov, V. ; Volknandt, Meiko ; Volkov, Vasily ; Neumann-Cosel, Peter von ; Schmid, Mirko von ; Wagner, A. ; Wamers, Felix ; Weick, Helmut ; Wells, D. ; Westerberg, L. ; Wieland, Oliver ; Wiescher, Michael ; Wimmer, Christine ; Wimmer, K. ; Winfield, John S. ; Winkel, M. ; Woods, Philip John ; Wyss, R. ; Yakorev, Dmitry ; Yavor, M. ; Zamora Cardona, Juan Carlos ; Zartova, I. ; Zerguerras, T. ; Zgura, M. ; Zhdanov, A. ; Zhukov, Mikhail V. ; Zieblinski, M. ; Zilges, Andreas ; Zuber, Kai
The nucleosynthesis of elements beyond iron is dominated by neutron captures in the s and r processes. However, 32 stable, proton-rich isotopes cannot be formed during those processes, because they are shielded from the s-process flow and r-process β-decay chains. These nuclei are attributed to the p and rp process.
For all those processes, current research in nuclear astrophysics addresses the need for more precise reaction data involving radioactive isotopes. Depending on the particular reaction, direct or inverse kinematics, forward or time-reversed direction are investigated to determine or at least to constrain the desired reaction cross sections.
The Facility for Antiproton and Ion Research (FAIR) will offer unique, unprecedented opportunities to investigate many of the important reactions. The high yield of radioactive isotopes, even far away from the valley of stability, allows the investigation of isotopes involved in processes as exotic as the r or rp processes.
The antibacterial properties of nanosilver have led to a versatile application spectrum including medical purposes and personal care products. However, the increasing use of nanosilver has raised concerns about its environmental impacts. Long-term exposure studies with aquatic invertebrates are essential to assess possible adverse effects on aquatic ecosystems. In the present study, acute (48 h), chronic (21 d) and long-term effects of nanosilver (primary size 15 nm) on five successive generations of three Daphnia species (D. magna, D. pulex, and D. galeata) were investigated. Acute EC50 values of nanosilver were 121 µg Ag L−1 for D. magna being the least sensitive species and 8.95 and 13.9 µg Ag L−1 for D. pulex and D. galeata, respectively. Chronic exposure provided EC10 values of 0.92 µg Ag L−1 for D. magna showing the most sensitive chronic reaction and 2.25 and 3.45 µg Ag L−1 for D. pulex and D. galeata, respectively. Comparative exposure to AgNO3 revealed a generally higher toxicity of the soluble form of silver. The multi-generation experiments resulted in effects on the population level for all tested species. Exposure of D. magna indicated an increased toxicity of nanosilver in the fifth generation of animals exposed to 10 µg Ag L−1. Neonates from pre-exposed parental daphnids did not completely recover when transferred into clean water. Exposure of D. pulex and D. galeata revealed not only increasing toxicity in some generations, but also greater tolerance to nanosilver. This study contributes to the assessment of the risk potential of nanosilver on aquatic ecosystems. It shows that effects of nanosilver vary within one genus and change with exposure duration. Therefore, long-term studies considering different aquatic species are needed to better understand the possible effects of nanosilver on aquatic ecosystems.
DNA damage in oocytes induces a switch of the quality control factor TAp63α from dimer to tetramer
(2011)
Deutsch, Gregor ; Zielonka, Elisabeth M. ; Coutandin, Daniel ; Weber, Tobias Alexander ; Schäfer, Birgit ; Hannewald, Jens ; Luh, Laura Martina ; Durst, Florian ; Ibrahim, Mohamed ; Hoffmann, Jan ; Niesen, Frank H. ; Sentürk, Aycan ; Kunkel, Hana ; Brutschy, Bernd ; Schleiff, Enrico ; Knapp, Stefan ; Acker-Palmer, Amparo ; Grez, Manuel ; McKeon, Frank ; Dötsch, Volker
TAp63a, a homolog of the p53 tumor suppressor, is a quality control factor in the female germline. Remarkably, already undamaged oocytes express high levels of the protein, suggesting that TAp63a’s activity is under tight control of an inhibitory mechanism. Biochemical studies have proposed that inhibition requires the C-terminal transactivation inhibitory domain. However, the structural mechanism of TAp63a inhibition remains unknown. Here, we show that TAp63a is kept in an inactive dimeric state. We reveal that relief of inhibition leads to tetramer formation with ~20-fold higher DNA affinity. In vivo, phosphorylation-triggered tetramerization of TAp63a is not reversible by dephosphorylation. Furthermore, we show that a helix in the oligomerization domain of p63 is crucial for tetramer stabilization and competes with the transactivation domain for the same binding site. Our results demonstrate how TAp63a is inhibited by complex domain-domain interactions that provide the basis for regulating quality control in oocytes.