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      High throughput techniques to reveal the molecular physiology and evolution of digestion in spiders

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          Abstract

          Background

          Spiders are known for their predatory efficiency and for their high capacity of digesting relatively large prey. They do this by combining both extracorporeal and intracellular digestion. Whereas many high throughput (“-omics”) techniques focus on biomolecules in spider venom, so far this approach has not yet been applied to investigate the protein composition of spider midgut diverticula (MD) and digestive fluid (DF).

          Results

          We here report on our investigations of both MD and DF of the spider Nephilingis ( Nephilengys) cruentata through the use of next generation sequencing and shotgun proteomics. This shows that the DF is composed of a variety of hydrolases including peptidases, carbohydrases, lipases and nuclease, as well as of toxins and regulatory proteins. We detect 25 astacins in the DF. Phylogenetic analysis of the corresponding transcript(s) in Arachnida suggests that astacins have acquired an unprecedented role for extracorporeal digestion in Araneae, with different orthologs used by each family. The results of a comparative study of spiders in distinct physiological conditions allow us to propose some digestion mechanisms in this interesting animal taxon.

          Conclusion

          All the high throughput data allowed the demonstration that DF is a secretion originating from the MD. We identified enzymes involved in the extracellular and intracellular phases of digestion. Besides that, data analyses show a large gene duplication event in Araneae digestive process evolution, mainly of astacin genes. We were also able to identify proteins expressed and translated in the digestive system, which until now had been exclusively associated to venom glands.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-016-3048-9) contains supplementary material, which is available to authorized users.

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          Most cited references33

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          Confidence Limits on Phylogenies: An Approach Using the Bootstrap

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            An alignment confidence score capturing robustness to guide tree uncertainty.

            Multiple sequence alignment (MSA) is the basis for a wide range of comparative sequence analyses from molecular phylogenetics to 3D structure prediction. Sophisticated algorithms have been developed for sequence alignment, but in practice, many errors can be expected and extensive portions of the MSA are unreliable. Hence, it is imperative to understand and characterize the various sources of errors in MSAs and to quantify site-specific alignment confidence. In this paper, we show that uncertainties in the guide tree used by progressive alignment methods are a major source of alignment uncertainty. We use this insight to develop a novel method for quantifying the robustness of each alignment column to guide tree uncertainty. We build on the widely used bootstrap method for perturbing the phylogenetic tree. Specifically, we generate a collection of trees and use each as a guide tree in the alignment algorithm, thus producing a set of MSAs. We next test the consistency of every column of the MSA obtained from the unperturbed guide tree with respect to the set of MSAs. We name this measure the "GUIDe tree based AligNment ConfidencE" (GUIDANCE) score. Using the Benchmark Alignment data BASE benchmark as well as simulation studies, we show that GUIDANCE scores accurately identify errors in MSAs. Additionally, we compare our results with the previously published Heads-or-Tails score and show that the GUIDANCE score is a better predictor of unreliably aligned regions.
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              Inferring duplications, losses, transfers and incomplete lineage sorting with nonbinary species trees

              Motivation: Gene duplication (D), transfer (T), loss (L) and incomplete lineage sorting (I) are crucial to the evolution of gene families and the emergence of novel functions. The history of these events can be inferred via comparison of gene and species trees, a process called reconciliation, yet current reconciliation algorithms model only a subset of these evolutionary processes. Results: We present an algorithm to reconcile a binary gene tree with a nonbinary species tree under a DTLI parsimony criterion. This is the first reconciliation algorithm to capture all four evolutionary processes driving tree incongruence and the first to reconcile non-binary species trees with a transfer model. Our algorithm infers all optimal solutions and reports complete, temporally feasible event histories, giving the gene and species lineages in which each event occurred. It is fixed-parameter tractable, with polytime complexity when the maximum species outdegree is fixed. Application of our algorithms to prokaryotic and eukaryotic data show that use of an incomplete event model has substantial impact on the events inferred and resulting biological conclusions. Availability: Our algorithms have been implemented in Notung, a freely available phylogenetic reconciliation software package, available at http://www.cs.cmu.edu/~durand/Notung. Contact: mstolzer@andrew.cmu.edu
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                Author and article information

                Contributors
                adriana.lopes@butantan.gov.br
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                7 September 2016
                7 September 2016
                2016
                : 17
                : 1
                : 716
                Affiliations
                [1 ]Laboratory of Biochemistry and Biophysics, Instituto Butantan, São Paulo, 05503-000 Brazil
                [2 ]Biotechnology Program, University of São Paulo, São Paulo, Brazil
                [3 ]Laboratory of Analytical Biotechnology and Innovative Peptide Biology, Delft University of Technology, Delft, The Netherlands
                [4 ]Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil
                [5 ]Department of Biology, University of Antwerp, Antwerp, Belgium
                Article
                3048
                10.1186/s12864-016-3048-9
                5013568
                26818753
                e23833e0-b6c9-4b47-8705-4854b47a0d58
                © The Author(s). 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 15 August 2015
                : 27 August 2016
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100003593, Conselho Nacional de Desenvolvimento Científico e Tecnológico (BR);
                Award ID: 237706/2012-1
                Award Recipient :
                Funded by: Netherlands proteomic centre
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                Genetics
                astacin,digestion,enzyme,high throughput (-omics) techniques,spider,arachnida,nephilingis (nephilengys cruentata)

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