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Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking
The Legionella pneumophila effector protein VipA is an actin nucleator that co‐localizes with actin filaments and early endosomes in infected macrophages and which interferes with organelle trafficking when expressed in yeast. To identify the regions of VipA involved in its subcellular localization...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4767423/ https://www.ncbi.nlm.nih.gov/pubmed/26626407 http://dx.doi.org/10.1002/mbo3.316 |
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author | Bugalhão, Joana N. Mota, Luís Jaime Franco, Irina S. |
author_facet | Bugalhão, Joana N. Mota, Luís Jaime Franco, Irina S. |
author_sort | Bugalhão, Joana N. |
collection | PubMed |
description | The Legionella pneumophila effector protein VipA is an actin nucleator that co‐localizes with actin filaments and early endosomes in infected macrophages and which interferes with organelle trafficking when expressed in yeast. To identify the regions of VipA involved in its subcellular localization and functions, we ectopically expressed specific VipA mutant proteins in eukaryotic cells. This indicated that the characteristic punctate distribution of VipA depends on its NH (2)‐terminal (amino acid residues 1–133) and central coiled‐coil (amino acid residues 133–206) regions, and suggested a role for the COOH‐terminal (amino acid residues 206–339) region in association with actin filaments and for the NH (2)‐terminal in co‐localization with early endosomes. Co‐immunoprecipitation and in vitro assays showed that the COOH‐terminal region of VipA is necessary and sufficient to mediate actin binding, and is essential but insufficient to induce microfilament formation. Assays in yeast revealed that the NH (2) and the COOH‐terminal regions, and possibly an NPY motif within the NH (2) region of VipA, are necessary for interference with organelle trafficking. Overall, this suggests that subversion of eukaryotic vesicular trafficking by VipA involves both its ability to associate with early endosomes via its NH (2)‐terminal region and its capacity to bind and polymerize actin through its COOH‐terminal region. |
format | Online Article Text |
id | pubmed-4767423 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-47674232016-03-07 Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking Bugalhão, Joana N. Mota, Luís Jaime Franco, Irina S. Microbiologyopen Original Research The Legionella pneumophila effector protein VipA is an actin nucleator that co‐localizes with actin filaments and early endosomes in infected macrophages and which interferes with organelle trafficking when expressed in yeast. To identify the regions of VipA involved in its subcellular localization and functions, we ectopically expressed specific VipA mutant proteins in eukaryotic cells. This indicated that the characteristic punctate distribution of VipA depends on its NH (2)‐terminal (amino acid residues 1–133) and central coiled‐coil (amino acid residues 133–206) regions, and suggested a role for the COOH‐terminal (amino acid residues 206–339) region in association with actin filaments and for the NH (2)‐terminal in co‐localization with early endosomes. Co‐immunoprecipitation and in vitro assays showed that the COOH‐terminal region of VipA is necessary and sufficient to mediate actin binding, and is essential but insufficient to induce microfilament formation. Assays in yeast revealed that the NH (2) and the COOH‐terminal regions, and possibly an NPY motif within the NH (2) region of VipA, are necessary for interference with organelle trafficking. Overall, this suggests that subversion of eukaryotic vesicular trafficking by VipA involves both its ability to associate with early endosomes via its NH (2)‐terminal region and its capacity to bind and polymerize actin through its COOH‐terminal region. John Wiley and Sons Inc. 2015-12-02 /pmc/articles/PMC4767423/ /pubmed/26626407 http://dx.doi.org/10.1002/mbo3.316 Text en © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Research Bugalhão, Joana N. Mota, Luís Jaime Franco, Irina S. Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title | Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title_full | Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title_fullStr | Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title_full_unstemmed | Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title_short | Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
title_sort | identification of regions within the legionella pneumophila vipa effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4767423/ https://www.ncbi.nlm.nih.gov/pubmed/26626407 http://dx.doi.org/10.1002/mbo3.316 |
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