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Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum
The highly conserved enzyme arginyl-tRNA-protein transferase (Ate1) mediates arginylation, a posttranslational modification that is only incompletely understood at its molecular level. To investigate whether arginylation affects actin-dependent processes in a simple model organism, Dictyostelium dis...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594445/ https://www.ncbi.nlm.nih.gov/pubmed/30586322 http://dx.doi.org/10.1091/mbc.E18-02-0132 |
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author | Batsios, Petros Ishikawa-Ankerhold, Hellen C. Roth, Heike Schleicher, Michael Wong, Catherine C. L. Müller-Taubenberger, Annette |
author_facet | Batsios, Petros Ishikawa-Ankerhold, Hellen C. Roth, Heike Schleicher, Michael Wong, Catherine C. L. Müller-Taubenberger, Annette |
author_sort | Batsios, Petros |
collection | PubMed |
description | The highly conserved enzyme arginyl-tRNA-protein transferase (Ate1) mediates arginylation, a posttranslational modification that is only incompletely understood at its molecular level. To investigate whether arginylation affects actin-dependent processes in a simple model organism, Dictyostelium discoideum, we knocked out the gene encoding Ate1 and characterized the phenotype of ate1-null cells. Visualization of actin cytoskeleton dynamics by live-cell microscopy indicated significant changes in comparison to wild-type cells. Ate1-null cells were almost completely lacking focal actin adhesion sites at the substrate-attached surface and were only weakly adhesive. In two-dimensional chemotaxis assays toward folate or cAMP, the motility of ate1-null cells was increased. However, in three-dimensional chemotaxis involving more confined conditions, the motility of ate1-null cells was significantly reduced. Live-cell imaging showed that GFP-tagged Ate1 rapidly relocates to sites of newly formed actin-rich protrusions. By mass spectrometric analysis, we identified four arginylation sites in the most abundant actin isoform of Dictyostelium, in addition to arginylation sites in other actin isoforms and several actin-binding proteins. In vitro polymerization assays with actin purified from ate1-null cells revealed a diminished polymerization capacity in comparison to wild-type actin. Our data indicate that arginylation plays a crucial role in the regulation of cytoskeletal activities. |
format | Online Article Text |
id | pubmed-6594445 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-65944452019-07-05 Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum Batsios, Petros Ishikawa-Ankerhold, Hellen C. Roth, Heike Schleicher, Michael Wong, Catherine C. L. Müller-Taubenberger, Annette Mol Biol Cell Articles The highly conserved enzyme arginyl-tRNA-protein transferase (Ate1) mediates arginylation, a posttranslational modification that is only incompletely understood at its molecular level. To investigate whether arginylation affects actin-dependent processes in a simple model organism, Dictyostelium discoideum, we knocked out the gene encoding Ate1 and characterized the phenotype of ate1-null cells. Visualization of actin cytoskeleton dynamics by live-cell microscopy indicated significant changes in comparison to wild-type cells. Ate1-null cells were almost completely lacking focal actin adhesion sites at the substrate-attached surface and were only weakly adhesive. In two-dimensional chemotaxis assays toward folate or cAMP, the motility of ate1-null cells was increased. However, in three-dimensional chemotaxis involving more confined conditions, the motility of ate1-null cells was significantly reduced. Live-cell imaging showed that GFP-tagged Ate1 rapidly relocates to sites of newly formed actin-rich protrusions. By mass spectrometric analysis, we identified four arginylation sites in the most abundant actin isoform of Dictyostelium, in addition to arginylation sites in other actin isoforms and several actin-binding proteins. In vitro polymerization assays with actin purified from ate1-null cells revealed a diminished polymerization capacity in comparison to wild-type actin. Our data indicate that arginylation plays a crucial role in the regulation of cytoskeletal activities. The American Society for Cell Biology 2019-02-15 /pmc/articles/PMC6594445/ /pubmed/30586322 http://dx.doi.org/10.1091/mbc.E18-02-0132 Text en © 2019 Batsios et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Articles Batsios, Petros Ishikawa-Ankerhold, Hellen C. Roth, Heike Schleicher, Michael Wong, Catherine C. L. Müller-Taubenberger, Annette Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title | Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title_full | Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title_fullStr | Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title_full_unstemmed | Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title_short | Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum |
title_sort | ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in dictyostelium discoideum |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594445/ https://www.ncbi.nlm.nih.gov/pubmed/30586322 http://dx.doi.org/10.1091/mbc.E18-02-0132 |
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