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      Investigation of potential pathogenicity of Willaertia magna by investigating the transfer of bacteria pathogenicity genes into its genome

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          Abstract

          Willaertia magna c2c maky is a thermophilic amoeba closely related to the genus Naegleria. This free-living amoeba has the ability to eliminate Legionella pneumophila, which is an amoeba-resisting bacterium living in an aquatic environment. To prevent the proliferation of L. pneumophila in cooling towers, the use of W. magna as natural biocide has been proposed. To provide a better understanding of the W. magna genome, whole-genome sequencing was performed through the study of virulence factors and lateral gene transfers. This amoeba harbors a genome of 36.5 megabases with 18,519 predicted genes. BLASTp analyses reported protein homology between 136  W. magna sequences and amoeba-resistant microorganisms. Horizontal gene transfers were observed based on the basis of the phylogenetic reconstruction hypothesis. We detected 15 homologs of N. fowleri genes related to virulence, although these latter were also found in the genome of N. gruberi, which is a non-pathogenic amoeba. Furthermore, the cytotoxicity test performed on human cells supports the hypothesis that the strain c2c maky is a non-pathogenic amoeba. This work explores the genomic repertory for the first draft genome of genus Willaertia and provides genomic data for further comparative studies on virulence of related pathogenic amoeba, N. fowleri.

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          The genome of the social amoeba Dictyostelium discoideum.

          The social amoebae are exceptional in their ability to alternate between unicellular and multicellular forms. Here we describe the genome of the best-studied member of this group, Dictyostelium discoideum. The gene-dense chromosomes of this organism encode approximately 12,500 predicted proteins, a high proportion of which have long, repetitive amino acid tracts. There are many genes for polyketide synthases and ABC transporters, suggesting an extensive secondary metabolism for producing and exporting small molecules. The genome is rich in complex repeats, one class of which is clustered and may serve as centromeres. Partial copies of the extrachromosomal ribosomal DNA (rDNA) element are found at the ends of each chromosome, suggesting a novel telomere structure and the use of a common mechanism to maintain both the rDNA and chromosomal termini. A proteome-based phylogeny shows that the amoebozoa diverged from the animal-fungal lineage after the plant-animal split, but Dictyostelium seems to have retained more of the diversity of the ancestral genome than have plants, animals or fungi.
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            Assessment of transcript reconstruction methods for RNA-seq

            RNA sequencing (RNA-seq) is transforming genome biology, enabling comprehensive transcriptome profiling with unprecendented accuracy and detail. Due to technical limitations of current high-throughput sequencing platforms, transcript identity, structure and expression level must be inferred programmatically from partial sequence reads of fragmented gene products. We evaluated 24 protocol variants of 14 independent computational methods for exon identification, transcript reconstruction and expression level quantification from RNA-seq data. Our results show that most algorithms are able to identify discrete transcript components with high success rates, but that assembly of complete isoform structures poses a major challenge even when all constituent elements are identified. Expression level estimates also varied widely across methods, even when based on similar transcript models. Consequently, the complexity of higher eukaryotic genomes imposes severe limitations in transcript recall and splice product discrimination that are likely to remain limiting factors for the analysis of current-generation RNA-seq data.
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              Microorganisms resistant to free-living amoebae.

              Free-living amoebae feed on bacteria, fungi, and algae. However, some microorganisms have evolved to become resistant to these protists. These amoeba-resistant microorganisms include established pathogens, such as Cryptococcus neoformans, Legionella spp., Chlamydophila pneumoniae, Mycobacterium avium, Listeria monocytogenes, Pseudomonas aeruginosa, and Francisella tularensis, and emerging pathogens, such as Bosea spp., Simkania negevensis, Parachlamydia acanthamoebae, and Legionella-like amoebal pathogens. Some of these amoeba-resistant bacteria (ARB) are lytic for their amoebal host, while others are considered endosymbionts, since a stable host-parasite ratio is maintained. Free-living amoebae represent an important reservoir of ARB and may, while encysted, protect the internalized bacteria from chlorine and other biocides. Free-living amoebae may act as a Trojan horse, bringing hidden ARB within the human "Troy," and may produce vesicles filled with ARB, increasing their transmission potential. Free-living amoebae may also play a role in the selection of virulence traits and in adaptation to survival in macrophages. Thus, intra-amoebal growth was found to enhance virulence, and similar mechanisms seem to be implicated in the survival of ARB in response to both amoebae and macrophages. Moreover, free-living amoebae represent a useful tool for the culture of some intracellular bacteria and new bacterial species that might be potential emerging pathogens.
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                Author and article information

                Contributors
                bernard.la-scola@univ-amu.fr
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                4 December 2019
                4 December 2019
                2019
                : 9
                Affiliations
                [1 ]ISNI 0000 0001 0407 1584, GRID grid.414336.7, Aix-Marseille Université UM63, Institut de Recherche pour le Développement IRD 198, Assistance Publique – Hôpitaux de Marseille (AP-HM), Microbes, Evolution, Phylogeny and Infection (MEΦI), Institut Hospitalo-Universitaire (IHU) - Méditerranée Infection, ; Marseille, France
                [2 ]Amoéba, Chassieu, France
                [3 ]ProfileXpert/Viroscan3D, UCBL UMS 3453 CNRS – US7 INSERM, Lyon, France
                [4 ]ISNI 0000 0004 0384 0005, GRID grid.462282.8, Inserm U1052, CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, ; Lyon, France
                [5 ]ISNI 0000 0001 2172 4233, GRID grid.25697.3f, Université de Lyon, ; Lyon, France
                Article
                54580
                10.1038/s41598-019-54580-6
                6892926
                31797948
                447f3ad4-e301-4d12-9b8d-1fdf6abe02fc
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001665, Agence Nationale de la Recherche (French National Research Agency);
                Award ID: Méditerranée Infection 10-IAHU-03
                Award ID: Méditerranée Infection 10-IAHU-03
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100008530, EC | European Regional Development Fund (Europski Fond za Regionalni Razvoj);
                Award ID: PRIMI
                Award Recipient :
                Categories
                Article
                Custom metadata
                © The Author(s) 2019

                Uncategorized
                computational biology and bioinformatics,phylogenetics
                Uncategorized
                computational biology and bioinformatics, phylogenetics

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