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      Molecular signatures that correlate with induction of lens regeneration in newts: lessons from proteomic analysis

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          Abstract

          Background

          Amphibians have the remarkable ability to regenerate missing body parts. After complete removal of the eye lens, the dorsal but not the ventral iris will transdifferentiate to regenerate an exact replica of the lost lens. We used reverse-phase nano-liquid chromatography followed by mass spectrometry to detect protein concentrations in dorsal and ventral iris 0, 4, and 8 days post-lentectomy. We performed gene expression comparisons between regeneration and intact timepoints as well as between dorsal and ventral iris.

          Results

          Our analysis revealed gene expression patterns associated with the ability of the dorsal iris for transdifferentiation and lens regeneration. Proteins regulating gene expression and various metabolic processes were enriched in regeneration timepoints. Proteins involved in extracellular matrix, gene expression, and DNA-associated functions like DNA repair formed a regeneration-related protein network and were all up-regulated in the dorsal iris. In addition, we investigated protein concentrations in cultured dorsal (transdifferentiation-competent) and ventral (transdifferentiation-incompetent) iris pigmented epithelial (IPE) cells. Our comparative analysis revealed that the ability of dorsal IPE cells to keep memory of their tissue of origin and transdifferentiation is associated with the expression of proteins that specify the dorso-ventral axis of the eye as well as with proteins found highly expressed in regeneration timepoints, especially 8 days post-lentectomy.

          Conclusions

          The study deepens our understanding in the mechanism of regeneration by providing protein networks and pathways that participate in the process.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s40246-014-0022-y) contains supplementary material, which is available to authorized users.

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          Most cited references52

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          Amputation-induced reactive oxygen species (ROS) are required for successful Xenopus tadpole tail regeneration

          Understanding the molecular mechanisms that promote successful tissue regeneration is critical for continued advancements in regenerative medicine. Vertebrate amphibian tadpoles of the species Xenopus laevis and Xenopus tropicalis have remarkable abilities to regenerate their tails following amputation 1, 2 , via the coordinated activity of numerous growth factor signaling pathways, including the Wnt, Fgf, BMP, notch, and TGFβ pathways 3-6 . Little is known, however, about the events that act upstream of these signalling pathways following injury. Here, we show that Xenopus tadpole tail amputation induces a sustained production of reactive oxygen species (ROS) during tail regeneration. Lowering ROS levels, via pharmacological or genetic approaches, reduces cell proliferation and impairs tail regeneration. Genetic rescue experiments restored both ROS production and the initiation of the regenerative response. Sustained increased ROS levels are required for Wnt/β-catenin signaling and the activation of one of its major downstream targets, fgf20 7 , which, in turn, is essential for proper tail regeneration. These findings demonstrate that injury-induced ROS production is an important regulator of tissue regeneration.
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            Kinase-selective enrichment enables quantitative phosphoproteomics of the kinome across the cell cycle.

            Protein kinases are pivotal regulators of cell signaling that modulate each other's functions and activities through site-specific phosphorylation events. These key regulatory modifications have not been studied comprehensively, because low cellular abundance of kinases has resulted in their underrepresentation in previous phosphoproteome studies. Here, we combine kinase-selective affinity purification with quantitative mass spectrometry to analyze the cell-cycle regulation of protein kinases. This proteomics approach enabled us to quantify 219 protein kinases from S and M phase-arrested human cancer cells. We identified more than 1000 phosphorylation sites on protein kinases. Intriguingly, half of all kinase phosphopeptides were upregulated in mitosis. Our data reveal numerous unknown M phase-induced phosphorylation sites on kinases with established mitotic functions. We also find potential phosphorylation networks involving many protein kinases not previously implicated in mitotic progression. These results provide a vastly extended knowledge base for functional studies on kinases and their regulation through site-specific phosphorylation.
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              MALAT-1 enhances cell motility of lung adenocarcinoma cells by influencing the expression of motility-related genes.

              MALAT-1, a long non-coding RNA, is associated with metastasis, but its role in the metastatic process remains unknown. Here, we show that short-interfering RNA-mediated MALAT-1 silencing impaired in vitro cell motility of lung cancer cells and influenced the expression of numerous genes. In these genes, knockdown of any one of CTHRC1, CCT4, HMMR, or ROD1 clearly inhibited cell migration. In MALAT-1 knockdown cells, pre-mRNA levels were decreased in some but not all genes. Thus, our findings suggest that MALAT-1 is a novel class of non-coding RNA that promotes cell motility through transcriptional and post-transcriptional regulation of motility related gene expression. Copyright © 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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                Author and article information

                Contributors
                sousounisk1@udayton.edu
                rbhavsar1@udayton.edu
                mario.looso@mpi-bn.mpg.de
                marcus.krueger@mpi-bn.mpg.de
                beebej1@udayton.edu
                thomas.braun@mpi-bn.mpg.de
                ptsonis1@udayton.edu
                Journal
                Hum Genomics
                Hum. Genomics
                Human Genomics
                BioMed Central (London )
                1473-9542
                1479-7364
                11 December 2014
                11 December 2014
                2014
                : 8
                : 1
                : 22
                Affiliations
                [ ]Department of Biology and Center for Tissue Regeneration and Engineering at Dayton, University of Dayton, 300 College Park, Dayton, OH 45469 USA
                [ ]Department of Cardiac Development and Remodeling, Max-Planck-Institute for Heart and Lung Research, Ludwigstrasse 43, 61231 Bad Nauheim, Germany
                Article
                22
                10.1186/s40246-014-0022-y
                4271507
                25496664
                35acca53-5c76-47f9-ae9a-e8c6b0208662
                © Sousounis et al.; licensee BioMed Central Ltd. 2014

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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
                : 3 November 2014
                : 17 November 2014
                Categories
                Primary Research
                Custom metadata
                © The Author(s) 2014

                Genetics
                regeneration,lens,newt,proteomics,gene expression,regeneration program
                Genetics
                regeneration, lens, newt, proteomics, gene expression, regeneration program

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