TY - INPR A1 - Hennig, Thomas A1 - Prusty, Archana B. A1 - Kaufer, Benedikt A1 - Whisnant, Adam W. A1 - Lodha, Manivel A1 - Enders, Antje A1 - Thomas, Julius A1 - Kasimir, Francesca A1 - Grothey, Arnhild A1 - Herb, Stefanie A1 - Jürges, Christopher A1 - Meister, Gunter A1 - Erhard, Florian A1 - Dölken, Lars A1 - Prusty, Bhupesh K. T1 - Selective inhibition of miRNA 1 processing by a herpesvirus encoded miRNA N2 - Herpesviruses have mastered host cell modulation and immune evasion to augment productive infection, life-long latency and reactivation thereof 1,2. A long appreciated, yet elusively defined relationship exists between the lytic-latent switch and viral non-coding RNAs 3,4. Here, we identify miRNA-mediated inhibition of miRNA processing as a thus far unknown cellular mechanism that human herpesvirus 6A (HHV-6A) exploits to disrupt mitochondrial architecture, evade intrinsic host defense and drive the lytic-latent switch. We demonstrate that virus-encoded miR-aU14 selectively inhibits the processing of multiple miR-30 family members by direct interaction with the respective pri-miRNA hairpin loops. Subsequent loss of miR-30 and activation of the miR-30/p53/Drp1 axis triggers a profound disruption of mitochondrial architecture. This impairs induction of type I interferons and is necessary for both productive infection and virus reactivation. Ectopic expression of miR-aU14 triggered virus reactivation from latency, identifying viral miR-aU14 as a readily drugable master regulator of the herpesvirus lytic-latent switch. Our results show that miRNA-mediated inhibition of miRNA processing represents a generalized cellular mechanism that can be exploited to selectively target individual members of miRNA families. We anticipate that targeting miR-aU14 provides exciting therapeutic options for preventing herpesvirus reactivations in HHV-6-associated disorders. KW - Herpesvirus KW - HHV-6A KW - miRNA processing KW - miR-30 KW - mitochondria KW - fusion and fission KW - type I interferon KW - latency KW - virus reactivation Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-267862 ET - accepted version ER - TY - INPR A1 - Hennig, Thomas A1 - Prusty, Archana B. A1 - Kaufer, Benedikt A1 - Whisnant, Adam W. A1 - Lodha, Manivel A1 - Enders, Antje A1 - Thomas, Julius A1 - Kasimir, Francesca A1 - Grothey, Arnhild A1 - Herb, Stefanie A1 - Jürges, Christopher A1 - Meister, Gunter A1 - Erhard, Florian A1 - Dölken, Lars A1 - Prusty, Bhupesh K. T1 - Selective inhibition of microRNA processing by a herpesvirus-encoded microRNA triggers virus reactivation from latency N2 - Herpesviruses have mastered host cell modulation and immune evasion to augment productive infection, life-long latency and reactivation thereof 1,2. A long appreciated, yet elusively defined relationship exists between the lytic-latent switch and viral non-coding RNAs 3,4. Here, we identify miRNA-mediated inhibition of miRNA processing as a novel cellular mechanism that human herpesvirus 6A (HHV-6A) exploits to disrupt mitochondrial architecture, evade intrinsic host defense and drive the latent-lytic switch. We demonstrate that virus-encoded miR-aU14 selectively inhibits the processing of multiple miR-30 family members by direct interaction with the respective pri-miRNA hairpin loops. Subsequent loss of miR-30 and activation of miR-30/p53/Drp1 axis triggers a profound disruption of mitochondrial architecture, which impairs induction of type I interferons and is necessary for both productive infection and virus reactivation. Ectopic expression of miR-aU14 was sufficient to trigger virus reactivation from latency thereby identifying it as a readily drugable master regulator of the herpesvirus latent-lytic switch. Our results show that miRNA-mediated inhibition of miRNA processing represents a generalized cellular mechanism that can be exploited to selectively target individual members of miRNA families. We anticipate that targeting miR-aU14 provides exciting therapeutic options for preventing herpesvirus reactivations in HHV-6-associated disorders like myalgic encephalitis/chronic fatigue syndrome (ME/CFS) and Long-COVID. KW - Herpesvirus KW - HHV-6 KW - miRNA processing KW - miR-30 KW - mitochondria KW - fusion and fission KW - type I interferon KW - latency KW - virus reactivation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-267858 UR - https://doi.org/10.21203/rs.3.rs-820696/v1 ET - submitted version ER - TY - JOUR A1 - Hendriksma, Harmen P. A1 - Küting, Meike A1 - Härtel, Stephan A1 - Näther, Astrid A1 - Dohrmann, Anja B. A1 - Steffan-Dewenter, Ingolf A1 - Tebbe, Christoph C. T1 - Effect of Stacked Insecticidal Cry Proteins from Maize Pollen on Nurse Bees (Apis mellifera carnica) and Their Gut Bacteria JF - PLoS ONE N2 - Honey bee pollination is a key ecosystem service to nature and agriculture. However, biosafety research on genetically modified crops rarely considers effects on nurse bees from intact colonies, even though they receive and primarily process the largest amount of pollen. The objective of this study was to analyze the response of nurse bees and their gut bacteria to pollen from Bt maize expressing three different insecticidal Cry proteins (Cry1A.105, Cry2Ab2, and Cry3Bb1). Naturally Cry proteins are produced by bacteria (Bacillus thuringiensis). Colonies of Apis mellifera carnica were kept during anthesis in flight cages on field plots with the Bt maize, two different conventionally bred maize varieties, and without cages, 1-km outside of the experimental maize field to allow ad libitum foraging to mixed pollen sources. During their 10-days life span, the consumption of Bt maize pollen had no effect on their survival rate, body weight and rates of pollen digestion compared to the conventional maize varieties. As indicated by ELISA-quantification of Cry1A.105 and Cry3Bb1, more than 98% of the recombinant proteins were degraded. Bacterial population sizes in the gut were not affected by the genetic modification. Bt-maize, conventional varieties and mixed pollen sources selected for significantly different bacterial communities which were, however, composed of the same dominant members, including Proteobacteria in the midgut and Lactobacillus sp. and Bifidobacterium sp. in the hindgut. Surprisingly, Cry proteins from natural sources, most likely B. thuringiensis, were detected in bees with no exposure to Bt maize. The natural occurrence of Cry proteins and the lack of detectable effects on nurse bees and their gut bacteria give no indication for harmful effects of this Bt maize on nurse honey bees. KW - communities KW - 16S ribosomal-RNA KW - T-RFLP analysis KW - honey bees KW - bacillus thuringiensis KW - risk assessment KW - multivariate analyses KW - worker honeybees KW - corn pollen KW - larvae Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-131025 VL - 8 IS - 3 ER - TY - JOUR A1 - Hendriksma, Harmen P. A1 - Härtel, Stephan A1 - Steffan-Dewenter, Ingolf T1 - Testing Pollen of Single and Stacked Insect-Resistant Bt-Maize on In vitro Reared Honey Bee Larvae JF - PLoS One N2 - The ecologically and economic important honey bee (Apis mellifera) is a key non-target arthropod species in environmental risk assessment (ERA) of genetically modified (GM) crops. Honey bee larvae are directly exposed to transgenic products by the consumption of GM pollen. But most ERA studies only consider responses of adult bees, although Bt-proteins primarily affect the larval phases of target organisms. We adopted an in vitro larvae rearing system, to assess lethal and sublethal effects of Bt-pollen consumption in a standardized eco-toxicological bioassay. The effects of pollen from two Bt-maize cultivars, one expressing a single and the other a total of three Bt-proteins, on the survival and prepupae weight of honey bee larvae were analyzed. The control treatments included pollen from three non-transgenic maize varieties and of Heliconia rostrata. Three days old larvae were fed the realistic exposure dose of 2 mg pollen within the semi-artificial diet. The larvae were monitored over 120 h, until the prepupal stage, where larvae terminate feeding and growing. Neither single nor stacked Bt-maize pollen showed an adverse effect on larval survival and the prepupal weight. In contrast, feeding of H. rostrata pollen caused significant toxic effects. The results of this study indicate that pollen of the tested Bt-varieties does not harm the development of in vitro reared A. mellifera larvae. To sustain the ecosystem service of pollination, Bt-impact on A. mellifera should always be a crucial part of regulatory biosafety assessments. We suggest that our approach of feeding GM pollen on in vitro reared honey bee larvae is well suited of becoming a standard bioassay in regulatory risk assessments schemes of GM crops. KW - larvae KW - pollen KW - insect pests KW - genetically modified plants KW - diet KW - genetically modified crops KW - maize KW - honey bees Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-137803 VL - 6 IS - 12 ER - TY - THES A1 - Hendriksma, Harmen P. T1 - Non-target effects of a multiple insect resistant Bt-maize on the honey bee (Apis mellifera L.) T1 - Nichtzieleffekte eines Bt-Mais mit multipler Insektenresistenz auf Honigbienen (Apis mellifera L.) N2 - Neue methodische Entwicklungen zur Untersuchung der Ursachen des weltweit beobachteten Bienensterbens sind nötig, um die lebenswichtige Ökosystemdienstleistung der Bestäubung zu gewährleisten. Die ökologisch und wirtschaftlich bedeutsame Honigbiene (Apis mellifera) ist ein wichtiger Nichtziel-Organismus im Zulassungsverfahren für gentechnisch veränderte Pflanzen. Bisher sind vor allem Methoden zur Testung erwachsener Bienen unter Laborbedingungen verwendet worden, aber für eine Risikobewertung mit Hilfe von standardisierten Bienenkolonien oder in vitro gezüchteten Honigbienenlarven sind keine robusten Methoden oder standardisierte Protokolle vorhanden. In dieser Arbeit wurde eine Vielzahl an neuen methodischen Ansätzen für die Biosicherheitsforschung entwickelt: eine Mortalitäts-Falle (Kapitel II), ein "Full-Life-Cycle" Test (III), eine robuste in vitro Aufzucht-Methodik (IV), ein standardisierter in vitro Test für Bt-Pollen (V), eine gemischte Toxizitätsprüfung für transgene Reinproteine (VI) und eine Überprüfung der Darmmikroflora sowie der Pollenverdauungrate (VII). Die Ergebnisse dieser Studien zeigten keine nachteiligen Wirkungen von Bt-Maispollen oder Bt-Reinproteinen im "Worst-Case" Szenario auf Honigbienen. In Anbetracht der Datenlage ist eine Schädigung der Honigbiene durch den getesteten Bt-Mais Mon89034xMon88017 unwahrscheinlich. Die Anwendung der Untersuchungsmethoden in zukünftigen Biosicherheitsstudien für transgene Pflanzen wird empfohlen. N2 - Honey bee pollination is an ecologically and economically important ecosystem service. New methodological developments are needed to research the underlying factors of globally observed bee losses. The honey bee (Apis mellifera) is a key non-target arthropod species for environmental risk assessment of genetically modified (GM) crops. For GM-crop risk assessments, mainly methods for monitoring adult honey bees under laboratory conditions are documented. However, protocols with robust methods for standardized colonies or in vitro reared honey bee larvae are currently lacking. Within the research, presented in this this dissertation, multiple methodological developments are achieved; a mortality trap (Chapter II), a ‘full life cycle test’ (III), a novel in vitro rearing methodology (IV), a standardized in vitro test for Bt-pollen (V), a mixed toxicity test for purified transgenic proteins (VI), and a bacterial flora test with pollen digestion rate monitoring (VII). Overall, the studies did not indicate a detrimental effect caused by Bt-maize pollen, or by purified Bt-proteins at worst case exposure levels. Considering the risk for honey bees and larvae, we conclude that the tested Bt-maize Mon89034xMon88017 is not likely to cause harm to honey bee colonies. The study methods presented are highly recommended for future environmental risk assessment studies testing GM-crop biosafety on honey bees. KW - Biene KW - Bt-Mais KW - Nichtzielorganismen KW - Bestäubung KW - Umwelttoxikologie KW - Honey bee KW - Apis mellifera KW - Environmental Risk Assessment KW - Bt-maize KW - Non-target effects KW - Öko-Toxikologie Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-70304 ER - TY - JOUR A1 - Hempelmann, Alexander A1 - Hartleb, Laura A1 - van Straaten, Monique A1 - Hashemi, Hamidreza A1 - Zeelen, Johan P. A1 - Bongers, Kevin A1 - Papavasiliou, F. Nina A1 - Engstler, Markus A1 - Stebbins, C. Erec A1 - Jones, Nicola G. T1 - Nanobody-mediated macromolecular crowding induces membrane fission and remodeling in the African trypanosome JF - Cell Reports N2 - The dense variant surface glycoprotein (VSG) coat of African trypanosomes represents the primary host-pathogen interface. Antigenic variation prevents clearing of the pathogen by employing a large repertoire of antigenically distinct VSG genes, thus neutralizing the host’s antibody response. To explore the epitope space of VSGs, we generate anti-VSG nanobodies and combine high-resolution structural analysis of VSG-nanobody complexes with binding assays on living cells, revealing that these camelid antibodies bind deeply inside the coat. One nanobody causes rapid loss of cellular motility, possibly due to blockage of VSG mobility on the coat, whose rapid endocytosis and exocytosis are mechanistically linked to Trypanosoma brucei propulsion and whose density is required for survival. Electron microscopy studies demonstrate that this loss of motility is accompanied by rapid formation and shedding of nanovesicles and nanotubes, suggesting that increased protein crowding on the dense membrane can be a driving force for membrane fission in living cells. KW - African trypanosome KW - host-pathogen interaction KW - variant surface glycoproteins KW - immune epitope mapping KW - structural biology KW - nanovesicle formation KW - nanotube formation KW - protein crowding KW - membrane fission Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270285 VL - 37 IS - 5 ER - TY - JOUR A1 - Helmreich, Ernst J. M. T1 - Ways and means of coping with uncertainties of the relationship of the genetic blue print to protein structure and function in the cell N2 - As one of the disciplines of systems biology, proteomics is central to enabling the elucidation of protein function within the cell; furthermore, the question of how to deduce protein structure and function from the genetic readout has gained new significance. This problem is of particular relevance for proteins engaged in cell signalling. In dealing with this question, I shall critically comment on the reliability and predictability of transmission and translation of the genetic blue print into the phenotype, the protein. Based on this information, I will then evaluate the intentions and goals of today’s proteomics and gene-networking and appraise their chances of success. Some of the themes commented on in this publication are explored in greater detail with particular emphasis on the historical roots of concepts and techniques in my forthcoming book, published in German: Von Molekülen zu Zellen. 100 Jahre experimentelle Biologie. Betrachtungen eines Biochemikers KW - Genetik Y1 - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-68006 ER - TY - JOUR A1 - Helmprobst, Frederik A1 - Lillesaar, Christina A1 - Stigloher, Christian T1 - Expression of sept3, sept5a and sept5b in the Developing and Adult Nervous System of the Zebrafish (Danio rerio) JF - Frontiers in Neuroanatomy N2 - Septins are a highly conserved family of small GTPases that form cytoskeletal filaments. Their cellular functions, especially in the nervous system, still remain largely enigmatic, but there are accumulating lines of evidence that septins play important roles in neuronal physiology and pathology. In order to further dissect septin function in the nervous system a detailed temporal resolved analysis in the genetically well tractable model vertebrate zebrafish (Danio rerio) is crucially necessary. To close this knowledge gap we here provide a reference dataset describing the expression of selected septins (sept3, sept5a and sept5b) in the zebrafish central nervous system. Strikingly, proliferation zones are devoid of expression of all three septins investigated, suggesting that they have a role in post-mitotic neural cells. Our finding that three septins are mainly expressed in non-proliferative regions was further confirmed by double-stainings with a proliferative marker. Our RNA in situ hybridization (ISH) study, detecting sept3, sept5a and sept5b mRNAs, shows that all three septins are expressed in largely overlapping regions of the developing brain. However, the expression of sept5a is much more confined compared to sept3 and sept5b. In contrast, the expression of all the three analyzed septins is largely similar in the adult brain. KW - retinal development KW - sept5b KW - septin KW - RNA in situ hybridization KW - neuronal development KW - sept3 KW - sept5a Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-157625 VL - 11 IS - 6 ER - TY - JOUR A1 - Helmprobst, Frederik A1 - Kneitz, Susanne A1 - Klotz, Barbara A1 - Naville, Magali A1 - Dechaud, Corentin A1 - Volff, Jean-Nicolas A1 - Schartl, Manfred T1 - Differential expression of transposable elements in the medaka melanoma model JF - PLoS One N2 - Malignant melanoma incidence is rising worldwide. Its treatment in an advanced state is difficult, and the prognosis of this severe disease is still very poor. One major source of these difficulties is the high rate of metastasis and increased genomic instability leading to a high mutation rate and the development of resistance against therapeutic approaches. Here we investigate as one source of genomic instability the contribution of activation of transposable elements (TEs) within the tumor. We used the well-established medaka melanoma model and RNA-sequencing to investigate the differential expression of TEs in wildtype and transgenic fish carrying melanoma. We constructed a medaka-specific TE sequence library and identified TE sequences that were specifically upregulated in tumors. Validation by qRT- PCR confirmed a specific upregulation of a LINE and an LTR element in malignant melanomas of transgenic fish. KW - melanoma KW - genomics KW - transposable elements KW - cancer genomics KW - malignant tumors KW - gene prediction KW - human genomics KW - retrotransposons Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260615 VL - 16 IS - 10 ER - TY - JOUR A1 - Heller, Klaus-Gerhard A1 - Hemp, Claudia T1 - Fiddler on the Tree - A Bush-Cricket Species with Unusual Stridulatory Organs and Song JF - PLOS ONE N2 - Insects of the order Orthoptera are well-known for their acoustic communication. The structures used for this purpose show a high diversity which obviously relates to differences in song parameters and to the physics of sound production. Here we describe song and morphology of the sound producing organs of a tropical bush-cricket, Ectomoptera nepicauda, from East Africa. It has a very unusual calling song consisting of frequency-modulated, pure-tone sounds in the high ultrasonic range of 80 to 120 kHz and produced by extremely fast wing movements. Concerning morphology, it represents the most extreme state in the degree of left-right fore-wing differentiation found among Orthoptera: the acoustic parts of the left fore-wing consist exclusively of the stridulatory file, comparable in function to the bow of a violin, while the right wing carries only the plectrum (= string) and mirror (= soundbox). KW - constraints KW - sound production KW - katydids orthoptera KW - mole crickets KW - tettigoniidae KW - mechanics Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117068 VL - 9 IS - 3 ER -