TY - JOUR A1 - Zou, Jie A1 - Wang, Weiwei A1 - Neffe, Axel T. A1 - Xu, Xun A1 - Li, Zhengdong A1 - Deng, Zijun A1 - Sun, Xianlei A1 - Ma, Nan A1 - Lendlein, Andreas T1 - Adipogenic differentiation of human adipose derived mesenchymal stem cells in 3D architectured gelatin based hydrogels (ArcGel) JF - Clinical hemorheology and microcirculation : blood flow and vessels N2 - Polymeric matrices mimicking multiple functions of the ECM are expected to enable a material induced regeneration of tissues. Here, we investigated the adipogenic differentiation of human adipose derived mesenchymal stem cells (hADSCs) in a 3D architectured gelatin based hydrogel (ArcGel) prepared from gelatin and L-lysine diisocyanate ethyl ester (LDI) in an one-step process, in which the formation of an open porous morphology and the chemical network formation were integrated. The ArcGel was designed to support adipose tissue regeneration with its 3D porous structure, high cell biocompatibility, and mechanical properties compatible with human subcutaneous adipose tissue. The ArcGel could support initial cell adhesion and survival of hADSCs. Under static culture condition, the cells could migrate into the inner part of the scaffold with a depth of 840 +/- 120 mu m after 4 days, and distributed in the whole scaffold (2mm in thickness) within 14 days. The cells proliferated in the scaffold and the fold increase of cell number after 7 days of culture was 2.55 +/- 0.08. The apoptotic rate of hADSCs in the scaffold was similar to that of cells maintained on tissue culture plates. When cultured in adipogenic induction medium, the hADSCs in the scaffold differentiated into adipocytes with a high efficiency (93 +/- 1%). Conclusively, this gelatin based 3D scaffold presented high cell compatibility for hADSC cultivation and differentiation, which could serve as a potential implant material in clinical applications for adipose tissue reparation and regeneration. KW - Mesenchymal stem cells KW - gelatin based scaffold KW - adipose tissue regeneration KW - adipogenic differentiation Y1 - 2017 U6 - https://doi.org/10.3233/CH-179210 SN - 1386-0291 SN - 1875-8622 VL - 67 IS - 3-4 SP - 297 EP - 307 PB - IOS Press CY - Amsterdam ER - TY - JOUR A1 - Zimmermann, Heike Hildegard A1 - Raschke, Elena A1 - Epp, Laura Saskia A1 - Stoof-Leichsenring, Kathleen Rosemarie A1 - Schirrmeister, Lutz A1 - Schwamborn, Georg A1 - Herzschuh, Ulrike T1 - The history of tree and shrub taxa on Bol'shoy Lyakhovsky Island (New Siberian Archipelago) since the Last Interglacial Uncovered by Sedimentary Ancient DNA and Pollen Data JF - Genes N2 - Ecosystem boundaries, such as the Arctic-Boreal treeline, are strongly coupled with climate and were spatially highly dynamic during past glacial-interglacial cycles. Only a few studies cover vegetation changes since the last interglacial, as most of the former landscapes are inundated and difficult to access. Using pollen analysis and sedimentary ancient DNA (sedaDNA) metabarcoding, we reveal vegetation changes on Bol’shoy Lyakhovsky Island since the last interglacial from permafrost sediments. Last interglacial samples depict high levels of floral diversity with the presence of trees (Larix, Picea, Populus) and shrubs (Alnus, Betula, Ribes, Cornus, Saliceae) on the currently treeless island. After the Last Glacial Maximum, Larix re-colonised the island but disappeared along with most shrub taxa. This was probably caused by Holocene sea-level rise, which led to increased oceanic conditions on the island. Additionally, we applied two newly developed larch-specific chloroplast markers to evaluate their potential for tracking past population dynamics from environmental samples. The novel markers were successfully re-sequenced and exhibited two variants of each marker in last interglacial samples. SedaDNA can track vegetation changes as well as genetic changes across geographic space through time and can improve our understanding of past processes that shape modern patterns. KW - sedaDNA KW - metabarcoding KW - trnL KW - single-nucleotide polymorphism (SNP) KW - treeline KW - MIS 5 to 1 KW - permafrost deposits KW - radiocarbon ages KW - palaeoenvironment KW - Larix Y1 - 2017 U6 - https://doi.org/10.3390/genes8100273 SN - 2073-4425 VL - 8 IS - 10 SP - 273 PB - MDPI CY - Basel ER - TY - THES A1 - Zimmermann, Heike Hildegard T1 - Vegetation changes and treeline dynamics in northern Siberia since the last interglacial revealed by sedimentary ancient DNA metabarcoding and organelle genome assembly of modern larches Y1 - 2017 ER - TY - JOUR A1 - Zibulski, Romy A1 - Wesener, Felix A1 - Wilkes, Heinz A1 - Plessen, Birgit A1 - Pestryakova, Luidmila Agafyevna A1 - Herzschuh, Ulrike T1 - C / N ratio, stable isotope (delta C-13, delta N-15), and n-alkane patterns of brown mosses along hydrological gradients of low-centred polygons of the Siberian Arctic JF - Biogeosciences N2 - Mosses are a major component of the arctic vegetation, particularly in wetlands. We present C / N atomic ratio, delta C-13 and delta N-15 data of 400 brown-moss samples belonging to 10 species that were collected along hydrological gradients within polygonal mires located on the southern Taymyr Peninsula and the Lena River delta in northern Siberia. Additionally, n-alkane patterns of six of these species (16 samples) were investigated. The aim of the study is to see whether the inter-and intraspecific differences in C / N, isotopic compositions and n-alkanes are indicative of habitat, particularly with respect to water level. Overall, we find high variability in all investigated parameters for two different moisture-related groups of moss species. The C / N ratios range between 11 and 53 (median: 32) and show large variations at the intraspecific level. However, species preferring a dry habitat (xero-mesophilic mosses) show higher C / N ratios than those preferring a wet habitat (meso-hygrophilic mosses). The delta C-13 values range between 37.0 and 22.5% (median D 27.8 %). The delta N-15 values range between 6.6 and C 1.7%(median D 2.2 %). We find differences in delta C-13 and delta N-15 compositions between both habitat types. For some species of the meso-hygrophilic group, we suggest that a relationship between the individ-ual habitat water level and isotopic composition can be inferred as a function of microbial symbiosis. The n-alkane distribution also shows differences primarily between xeromesophilic and meso-hygrophilic mosses, i. e. having a dominance of n-alkanes with long (n-C29, n-C31 /and intermediate (n-C25 /chain lengths, respectively. Overall, our results reveal that C / N ratios, isotopic signals and n-alkanes of studied brown-moss taxa from polygonal wetlands are characteristic of their habitat. Y1 - 2017 U6 - https://doi.org/10.5194/bg-14-1617-2017 SN - 1726-4170 SN - 1726-4189 VL - 14 SP - 1617 EP - 1630 PB - Copernicus CY - Göttingen ER - TY - JOUR A1 - Zhu, Fangjun A1 - Schlupp, Ingo A1 - Tiedemann, Ralph T1 - Allele-specific expression at the androgen receptor alpha gene in a hybrid unisexual fish, the Amazon molly (Poecilia formosa) JF - PLoS one N2 - The all-female Amazon molly (Poecilia formosa) is the result of a hybridization of the Atlantic molly (P. mexicana) and the sailfin molly (P. latipinna) approximately 120,000 years ago. As a gynogenetic species, P. formosa needs to copulate with heterospecific males including males from one of its bisexual ancestral species. However, the sperm only triggers embryogenesis of the diploid eggs. The genetic information of the sperm donor typically will not contribute to the next generation of P. formosa. Hence, P. formosa possesses generally one allele from each of its ancestral species at any genetic locus. This raises the question whether both ancestral alleles are equally expressed in P. formosa. Allele-specific expression (ASE) has been previously assessed in various organisms, e.g., human and fish, and ASE was found to be important in the context of phenotypic variability and disease. In this study, we utilized Real-Time PCR techniques to estimate ASE of the androgen receptor alpha (arα) gene in several distinct tissues of Amazon mollies. We found an allelic bias favoring the maternal ancestor (P. mexicana) allele in ovarian tissue. This allelic bias was not observed in the gill or the brain tissue. Sequencing of the promoter regions of both alleles revealed an association between an Indel in a known CpG island and differential expression. Future studies may reveal whether our observed cis-regulatory divergence is caused by an ovary-specific trans-regulatory element, preferentially activating the allele of the maternal ancestor. Y1 - 2017 U6 - https://doi.org/10.1371/journal.pone.0186411 SN - 1932-6203 VL - 12 IS - 10 SP - 1 EP - 14 PB - PLoS CY - Lawrence, Kan. ER - TY - GEN A1 - Zhang, Yunming A1 - Lenhard, Michael T1 - Exiting Already? Molecular Control of Cell-Proliferation Arrest in Leaves: Cutting Edge T2 - Molecular plant Y1 - 2017 U6 - https://doi.org/10.1016/j.molp.2017.05.004 SN - 1674-2052 SN - 1752-9867 VL - 10 SP - 909 EP - 911 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Zeng, Ting A1 - Leimkühler, Silke A1 - Wollenberger, Ulla A1 - Fourmond, Vincent T1 - Transient Catalytic Voltammetry of Sulfite Oxidase Reveals Rate Limiting Conformational Changes JF - Journal of the American Chemical Society N2 - Sulfite oxidases are metalloenzymes that oxidize sulfite to sulfate at a molybdenum active site. In vertebrate sulfite oxidases, the electrons generated at the Mo center are transferred to an external electron acceptor via a heme domain, which can adopt two conformations: a “closed” conformation, suitable for internal electron transfer, and an “open” conformation suitable for intermolecular electron transfer. This conformational change is an integral part of the catalytic cycle. Sulfite oxidases have been wired to electrode surfaces, but their immobilization leads to a significant decrease in their catalytic activity, raising the question of the occurrence of the conformational change when the enzyme is on an electrode. We recorded and quantitatively modeled for the first time the transient response of the catalytic cycle of human sulfite oxidase immobilized on an electrode. We show that conformational changes still occur on the electrode, but at a lower rate than in solution, which is the reason for the decrease in activity of sulfite oxidases upon immobilization. Y1 - 2017 U6 - https://doi.org/10.1021/jacs.7b05480 SN - 0002-7863 VL - 139 SP - 11559 EP - 11567 PB - American Chemical Society CY - Washington ER - TY - THES A1 - Zeng, Ting T1 - Nanoparticles promoted biocatalysis BT - Electrochemical investigation of human sulfite oxidase on nanoparticles modified electrodes Y1 - 2017 ER - TY - JOUR A1 - Yarman, Aysu A1 - Jetzschmann, Katharina J. A1 - Neumann, Bettina A1 - Zhang, Xiaorong A1 - Wollenberger, Ulla A1 - Cordin, Aude A1 - Haupt, Karsten A1 - Scheller, Frieder W. T1 - Enzymes as Tools in MIP-Sensors JF - Chemosensors N2 - Molecularly imprinted polymers (MIPs) have the potential to complement antibodies in bioanalysis, are more stable under harsh conditions, and are potentially cheaper to produce. However, the affinity and especially the selectivity of MIPs are in general lower than those of their biological pendants. Enzymes are useful tools for the preparation of MIPs for both low and high-molecular weight targets: As a green alternative to the well-established methods of chemical polymerization, enzyme-initiated polymerization has been introduced and the removal of protein templates by proteases has been successfully applied. Furthermore, MIPs have been coupled with enzymes in order to enhance the analytical performance of biomimetic sensors: Enzymes have been used in MIP-sensors as tracers for the generation and amplification of the measuring signal. In addition, enzymatic pretreatment of an analyte can extend the analyte spectrum and eliminate interferences. KW - enzymatic MIP synthesis KW - template digestion KW - enzyme tracer KW - enzymatic analyte conversion KW - molecularly imprinted polymers Y1 - 2017 U6 - https://doi.org/10.3390/chemosensors5020011 SN - 2227-9040 VL - 5 PB - MDPI CY - Basel ER - TY - JOUR A1 - Yarman, Aysu T1 - Development of a molecularly imprinted polymer-based electrochemical sensor for tyrosinase JF - Turkish journal of chemistry N2 - For the first time a molecularly imprinted polymer (MIP)-based sensor for tyrosinase is described. This sensor is based on the electropolymerization of scopoletin or o-phenylenediamine in the presence of tyrosinase from mushrooms, which has a high homology to the human enzyme. The template was removed either by treatment with proteinase Kor by alkaline treatment. The measuring signal was generated either by measuring the formation of a product by the target enzyme or by evaluation of the permeability of the redox marker ferricyanide. The o-phenylenediamine-based MIP sensor has a linear measuring range up to 50 nM of tyrosinase with a limit of detection of 3.97 nM (R 2 = 0.994) and shows good discrimination towards other proteins, e.g., bovine serum albumin and cytochrome c. KW - Molecularly imprinted polymers KW - biomimetic sensors KW - tyrosinase KW - electropolymerization KW - scopoletin KW - ophenylenediamine Y1 - 2017 U6 - https://doi.org/10.3906/kim-1708-68 SN - 1300-0527 VL - 42 IS - 2 SP - 346 EP - 354 PB - Türkiye Bilimsel ve Teknik Araştırma Kurumu CY - Ankara ER - TY - THES A1 - Yang, Lei T1 - Verification of systemic mRNAs mobility and mobile functions Y1 - 2017 ER - TY - JOUR A1 - Wurzbacher, Christian A1 - Fuchs, Andrea A1 - Attermeyer, Katrin A1 - Frindte, Katharina A1 - Grossart, Hans-Peter A1 - Hupfer, Michael A1 - Casper, Peter A1 - Monaghan, Michael T. T1 - Shifts among Eukaryota, Bacteria, and Archaea define the vertical organization of a lake sediment JF - Microbiome N2 - Background: Lake sediments harbor diverse microbial communities that cycle carbon and nutrients while being constantly colonized and potentially buried by organic matter sinking from the water column. The interaction of activity and burial remained largely unexplored in aquatic sediments. We aimed to relate taxonomic composition to sediment biogeochemical parameters, test whether community turnover with depth resulted from taxonomic replacement or from richness effects, and to provide a basic model for the vertical community structure in sediments. Methods: We analyzed four replicate sediment cores taken from 30-m depth in oligo-mesotrophic Lake Stechlin in northern Germany. Each 30-cm core spanned ca. 170 years of sediment accumulation according to Cs-137 dating and was sectioned into layers 1-4 cm thick. We examined a full suite of biogeochemical parameters and used DNA metabarcoding to examine community composition of microbial Archaea, Bacteria, and Eukaryota. Results: Community beta-diversity indicated nearly complete turnover within the uppermost 30 cm. We observed a pronounced shift from Eukaryota- and Bacteria-dominated upper layers (<5 cm) to Bacteria-dominated intermediate layers (5-14 cm) and to deep layers (>14 cm) dominated by enigmatic Archaea that typically occur in deep-sea sediments. Taxonomic replacement was the prevalent mechanism in structuring the community composition and was linked to parameters indicative of microbial activity (e.g., CO2 and CH4 concentration, bacterial protein production). Richness loss played a lesser role but was linked to conservative parameters (e.g., C, N, P) indicative of past conditions. Conclusions: By including all three domains, we were able to directly link the exponential decay of eukaryotes with the active sediment microbial community. The dominance of Archaea in deeper layers confirms earlier findings from marine systems and establishes freshwater sediments as a potential low-energy environment, similar to deep sea sediments. We propose a general model of sediment structure and function based on microbial characteristics and burial processes. An upper "replacement horizon" is dominated by rapid taxonomic turnover with depth, high microbial activity, and biotic interactions. A lower "depauperate horizon" is characterized by low taxonomic richness, more stable "low-energy" conditions, and a dominance of enigmatic Archaea. KW - Archaea KW - Eukaryota KW - Bacteria KW - Community KW - Freshwater KW - Lake KW - DNA metabarcoding KW - Beta-diversity KW - Sediment KW - Turnover Y1 - 2017 U6 - https://doi.org/10.1186/s40168-017-0255-9 SN - 2049-2618 VL - 5 PB - BioMed Central CY - London ER - TY - JOUR A1 - Wurzbacher, Christian A1 - Attermeyer, Katrin A1 - Kettner, Marie Therese A1 - Flintrop, Clara A1 - Warthmann, Norman A1 - Hilt, Sabine A1 - Grossart, Hans-Peter A1 - Monaghan, Michael T. T1 - DNA metabarcoding of unfractionated water samples relates phyto-, zoo- and bacterioplankton dynamics and reveals a single-taxon bacterial bloom JF - Environmental microbiology reports N2 - Most studies of aquatic plankton focus on either macroscopic or microbial communities, and on either eukaryotes or prokaryotes. This separation is primarily for methodological reasons, but can overlook potential interactions among groups. Here we tested whether DNA metabarcoding of unfractionated water samples with universal primers could be used to qualitatively and quantitatively study the temporal dynamics of the total plankton community in a shallow temperate lake. Significant changes in the relative proportions of normalized sequence reads of eukaryotic and prokaryotic plankton communities over a 3-month period in spring were found. Patterns followed the same trend as plankton estimates measured using traditional microscopic methods. The bloom of a conditionally rare bacterial taxon belonging to Arcicella was characterized, which rapidly came to dominate the whole lake ecosystem and would have remained unnoticed without metabarcoding. The data demonstrate the potential of universal DNA metabarcoding applied to unfractionated samples for providing a more holistic view of plankton communities. Y1 - 2017 U6 - https://doi.org/10.1111/1758-2229.12540 SN - 1758-2229 VL - 9 SP - 383 EP - 388 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Wu, Yabin A1 - Stoddart, Martin J. A1 - Wuertz-Kozak, Karin A1 - Grad, Sibylle A1 - Alini, Mauro A1 - Ferguson, Stephen J. T1 - Hyaluronan supplementation as a mechanical regulator of cartilage tissue development under joint-kinematic-mimicking loading JF - Interface : journal of the Royal Society KW - synovial fluid KW - cartilage tissue engineering KW - mechanical loading KW - non-Newtonian fluid KW - hyaluronic acid supplementation Y1 - 2017 U6 - https://doi.org/10.1098/rsif.2017.0255 SN - 1742-5689 SN - 1742-5662 VL - 14 PB - Royal Society CY - London ER - TY - THES A1 - Wu, Si T1 - Exploring the Arabidopsis metabolic landscape by genetic mapping integrated with network analysis Y1 - 2017 ER - TY - JOUR A1 - Wolff, Martin A1 - Zhang-Haagen, Bo A1 - Decker, Christina A1 - Barz, Bogdan A1 - Schneider, Mario A1 - Biehl, Ralf A1 - Radulescu, Aurel A1 - Strodel, Birgit A1 - Willbold, Dieter A1 - Nagel-Steger, Luitgard T1 - A beta 42 pentamers/hexamers are the smallest detectable oligomers in solution JF - Scientific reports Y1 - 2017 U6 - https://doi.org/10.1038/s41598-017-02370-3 SN - 2045-2322 VL - 7 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Witt, B. A1 - Bornhorst, Julia A1 - Mitze, H. A1 - Ebert, Franziska A1 - Meyer, S. A1 - Francesconi, Kevin A. A1 - Schwerdtle, Tanja T1 - Arsenolipids exert less toxicity in a human neuron astrocyte co-culture as compared to the respective monocultures JF - Metallomics : integrated biometal science N2 - Arsenic-containing hydrocarbons (AsHCs), natural products found in seafood, have recently been shown to exert toxic effects in human neurons. In this study we assessed the toxicity of three AsHCs in cultured human astrocytes. Due to the high cellular accessibility and substantial toxicity observed astrocytes were identified as further potential brain target cells for arsenolipids. Thereby, the AsHCs exerted a 5-19-fold higher cytotoxicity in astrocytes as compared to arsenite. Next we compared the toxicity of the arsenicals in a co-culture model of the respective human astrocytes and neurons. Notably the AsHCs did not show any substantial toxic effects in the co-culture, while arsenite did. The arsenic accessibility studies indicated that in the co-culture astrocytes protect neurons against cellular arsenic accumulation especially after incubation with arsenolipids. In summary, these data underline the importance of the glial-neuron interaction when assessing the in vitro neurotoxicity of new unclassified metal species. Y1 - 2017 U6 - https://doi.org/10.1039/c7mt00036g SN - 1756-5901 SN - 1756-591X VL - 9 SP - 442 EP - 446 PB - Royal Society of Chemistry CY - Cambridge ER - TY - JOUR A1 - Wiegmann, Alex A1 - Rutschmann, Ronja A1 - Willemsen, Pascale T1 - Empirically investigating the concept of lying JF - Journal of Indian Council of Philosophical Research N2 - Lying is an everyday moral phenomenon about which philosophers have written a lot. Not only the moral status of lying has been intensively discussed but also what it means to lie in the first place. Perhaps the most important criterion for an adequate definition of lying is that it fits with people’s understanding and use of this concept. In this light, it comes as a surprise that researchers only recently started to empirically investigate the folk concept of lying. In this paper, we describe three experimental studies which address the following questions: Does a statement need to be objectively false in order to constitute lying? Does lying necessarily include the intention to deceive? Can one lie by omitting relevant facts? KW - Concept of lying KW - Lying KW - Definition of lying KW - Experimental philosophy Y1 - 2017 U6 - https://doi.org/10.1007/s40961-017-0112-z SN - 0970-7794 SN - 2363-9962 VL - 34 SP - 591 EP - 609 PB - Springer CY - New Dehli ER - TY - JOUR A1 - Wieczorek, Mareike A1 - Kruse, Stefan A1 - Epp, Laura Saskia A1 - Kolmogorov, Alexei A1 - Nikolaev, Anatoly N. A1 - Heinrich, Ingo A1 - Jeltsch, Florian A1 - Pestryakova, Luidmila Agafyevna A1 - Zibulski, Romy A1 - Herzschuh, Ulrike T1 - Dissimilar responses of larch stands in northern Siberia to increasing temperatures-a field and simulation based study JF - Ecology : a publication of the Ecological Society of America N2 - Arctic and alpine treelines worldwide differ in their reactions to climate change. A northward advance of or densification within the treeline ecotone will likely influence climate-vegetation feedback mechanisms. In our study, which was conducted in the Taimyr Depression in the North Siberian Lowlands, w present a combined field-and model-based approach helping us to better understand the population processes involved in the responses of the whole treeline ecotone, spanning from closed forest to single-tree tundra, to climate warming. Using information on stand structure, tree age, and seed quality and quantity from seven sites, we investigate effects of intra-specific competition and seed availability on the specific impact of recent climate warming on larch stands. Field data show that tree density is highest in the forest-tundra, and average tree size decreases from closed forest to single-tree tundra. Age-structure analyses indicate that the trees in the closed forest and forest-tundra have been present for at least similar to 240 yr. At all sites except the most southerly ones, past establishment is positively correlated with regional temperature increase. In the single-tree tundra, however, a change in growth form from krummholz to erect trees, beginning similar to 130 yr ago, rather than establishment date has been recorded. Seed mass decreases from south to north, while seed quantity increases. Simulations with LAVESI (Larix Vegetation Simulator) further suggest that relative density changes strongly in response to a warming signal in the forest-tundra while intra-specific competition limits densification in the closed forest and seed limitation hinders densification in the single-tree tundra. We find striking differences in strength and timing of responses to recent climate warming. While forest-tundra stands recently densified, recruitment is almost non-existent at the southern and northern end of the ecotone due to autecological processes. Palaeo-treelines may therefore be inappropriate to infer past temperature changes at a fine scale. Moreover, a lagged treeline response to past warming will, via feedback mechanisms, influence climate change in the future. KW - climate change KW - closed forest KW - dendroecology KW - forest change KW - latitude KW - recruitment KW - tundra KW - vegetation model Y1 - 2017 U6 - https://doi.org/10.1002/ecy.1887 SN - 0012-9658 SN - 1939-9170 VL - 98 SP - 2343 EP - 2355 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Whiteley, Liam A1 - Haug, Maria A1 - Klein, Kristina A1 - Willmann, Matthias A1 - Bohn, Erwin A1 - Chiantia, Salvatore A1 - Schwarz, Sandra T1 - Cholesterol and host cell surface proteins contribute to cell-cell fusion induced by the Burkholderia type VI secretion system 5 JF - PLoS one N2 - Following escape into the cytoplasm of host cells, Burkholderia pseudomallei and the related species Burkholderia thailandensis employ the type VI secretion system 5 ( T6SS-5) to induce plasma membrane fusion with an adjacent host cell. This process leads to the formation of multinucleated giant cells and facilitates bacterial access to an uninfected host cell in a direct manner. Despite its importance in virulence, the mechanism of the T6SS-5 and the role of host cell factors in cell-cell fusion remain elusive. To date, the T6SS-5 is the only system of bacterial origin known to induce host-cell fusion. To gain insight into the nature of T6SS-5-stimulated membrane fusion, we investigated the contribution of cholesterol and proteins exposed on the host cell surface, which were shown to be critically involved in virus-mediated giant cell formation. In particular, we analyzed the effect of host cell surface protein and cholesterol depletion on the formation of multinucleated giant cells induced by B. thailandensis. Acute protease treatment of RAW264.7 macrophages during infection with B. thailandensis followed by agarose overlay assays revealed a strong reduction in the number of cell-cell fusions compared with EDTA treated cells. Similarly, proteolytic treatment of specifically infected donor cells or uninfected recipient cells significantly decreased multinucleated giant cell formation. Furthermore, modulating host cell cholesterol content by acute cholesterol depletion from cellular membranes by methyl-beta-cyclodextrin treatment or exogenous addition of cholesterol impaired the ability of B. thailandensis to induce cell-cell fusions. The requirement of physiological cholesterol levels suggests that the membrane organization or mechanical properties of the lipid bilayer influence the fusion process. Altogether, our data suggest that membrane fusion induced by B. pseudomallei and B. thailandensis involves a complex interplay between the T6SS-5 and the host cell. Y1 - 2017 U6 - https://doi.org/10.1371/journal.pone.0185715 SN - 1932-6203 VL - 12 PB - PLoS CY - San Fransisco ER -