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Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity

Leishmania parasites are thought to control protein activity at the post-translational level, e.g. by protein phosphorylation. In the pathogenic amastigote, the mammalian stage of Leishmania parasites, heat shock proteins show increased phosphorylation, indicating a role in stage-specific signal tra...

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Autores principales: Hombach-Barrigah, Antje, Bartsch, Katharina, Smirlis, Despina, Rosenqvist, Heidi, MacDonald, Andrea, Dingli, Florent, Loew, Damarys, Späth, Gerald F., Rachidi, Najma, Wiese, Martin, Clos, Joachim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434042/
https://www.ncbi.nlm.nih.gov/pubmed/30911045
http://dx.doi.org/10.1038/s41598-019-41640-0
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author Hombach-Barrigah, Antje
Bartsch, Katharina
Smirlis, Despina
Rosenqvist, Heidi
MacDonald, Andrea
Dingli, Florent
Loew, Damarys
Späth, Gerald F.
Rachidi, Najma
Wiese, Martin
Clos, Joachim
author_facet Hombach-Barrigah, Antje
Bartsch, Katharina
Smirlis, Despina
Rosenqvist, Heidi
MacDonald, Andrea
Dingli, Florent
Loew, Damarys
Späth, Gerald F.
Rachidi, Najma
Wiese, Martin
Clos, Joachim
author_sort Hombach-Barrigah, Antje
collection PubMed
description Leishmania parasites are thought to control protein activity at the post-translational level, e.g. by protein phosphorylation. In the pathogenic amastigote, the mammalian stage of Leishmania parasites, heat shock proteins show increased phosphorylation, indicating a role in stage-specific signal transduction. Here we investigate the impact of phosphosites in the L. donovani heat shock protein 90. Using a chemical knock-down/genetic complementation approach, we mutated 11 confirmed or presumed phosphorylation sites and assessed the impact on overall fitness, morphology and in vitro infectivity. Most phosphosite mutations affected the growth and morphology of promastigotes in vitro, but with one exception, none of the phosphorylation site mutants had a selective impact on the in vitro infection of macrophages. Surprisingly, aspartate replacements mimicking the negative charge of phosphorylated serines or threonines had mostly negative impacts on viability and infectivity. HSP90 is a substrate for casein kinase 1.2-catalysed phosphorylation in vitro. While several putative phosphosite mutations abrogated casein kinase 1.2 activity on HSP90, only Ser(289) could be identified as casein kinase target by mass spectrometry. In summary, our data show HSP90 as a downstream client of phosphorylation-mediated signalling in an organism that depends on post-transcriptional gene regulation.
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spelling pubmed-64340422019-04-02 Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity Hombach-Barrigah, Antje Bartsch, Katharina Smirlis, Despina Rosenqvist, Heidi MacDonald, Andrea Dingli, Florent Loew, Damarys Späth, Gerald F. Rachidi, Najma Wiese, Martin Clos, Joachim Sci Rep Article Leishmania parasites are thought to control protein activity at the post-translational level, e.g. by protein phosphorylation. In the pathogenic amastigote, the mammalian stage of Leishmania parasites, heat shock proteins show increased phosphorylation, indicating a role in stage-specific signal transduction. Here we investigate the impact of phosphosites in the L. donovani heat shock protein 90. Using a chemical knock-down/genetic complementation approach, we mutated 11 confirmed or presumed phosphorylation sites and assessed the impact on overall fitness, morphology and in vitro infectivity. Most phosphosite mutations affected the growth and morphology of promastigotes in vitro, but with one exception, none of the phosphorylation site mutants had a selective impact on the in vitro infection of macrophages. Surprisingly, aspartate replacements mimicking the negative charge of phosphorylated serines or threonines had mostly negative impacts on viability and infectivity. HSP90 is a substrate for casein kinase 1.2-catalysed phosphorylation in vitro. While several putative phosphosite mutations abrogated casein kinase 1.2 activity on HSP90, only Ser(289) could be identified as casein kinase target by mass spectrometry. In summary, our data show HSP90 as a downstream client of phosphorylation-mediated signalling in an organism that depends on post-transcriptional gene regulation. Nature Publishing Group UK 2019-03-25 /pmc/articles/PMC6434042/ /pubmed/30911045 http://dx.doi.org/10.1038/s41598-019-41640-0 Text en © 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/.
spellingShingle Article
Hombach-Barrigah, Antje
Bartsch, Katharina
Smirlis, Despina
Rosenqvist, Heidi
MacDonald, Andrea
Dingli, Florent
Loew, Damarys
Späth, Gerald F.
Rachidi, Najma
Wiese, Martin
Clos, Joachim
Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title_full Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title_fullStr Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title_full_unstemmed Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title_short Leishmania donovani 90 kD Heat Shock Protein – Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity
title_sort leishmania donovani 90 kd heat shock protein – impact of phosphosites on parasite fitness, infectivity and casein kinase affinity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434042/
https://www.ncbi.nlm.nih.gov/pubmed/30911045
http://dx.doi.org/10.1038/s41598-019-41640-0
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