Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>

ABSTRACT The 90-kDa heat shock protein (HSP90) of eukaryotes is a highly abundant and essential chaperone required for the maturation of regulatory and signal proteins. In the protozoan parasite Leishmania donovani, causative agent of the fatal visceral leishmaniasis, HSP90 activity is essential for...

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Autores principales:	Eugenia Bifeld, Stephan Lorenzen, Katharina Bartsch, Juan-José Vasquez, T. Nicolai Siegel, Joachim Clos
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	HSP90 Leishmania donovani radicicol ribosome profiling Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5dbeae5ae8294a028155e3a33ec51390
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spelling	oai:doaj.org-article:5dbeae5ae8294a028155e3a33ec513902021-12-02T19:46:17ZRibosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>10.1128/mSystems.00214-182379-5077https://doaj.org/article/5dbeae5ae8294a028155e3a33ec513902018-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00214-18https://doaj.org/toc/2379-5077ABSTRACT The 90-kDa heat shock protein (HSP90) of eukaryotes is a highly abundant and essential chaperone required for the maturation of regulatory and signal proteins. In the protozoan parasite Leishmania donovani, causative agent of the fatal visceral leishmaniasis, HSP90 activity is essential for cell proliferation and survival. Even more importantly, its inhibition causes life cycle progression from the insect stage to the pathogenic, mammalian stage. To unravel the molecular impact of HSP90 activity on the parasites’ gene expression, we performed a ribosome profiling analysis of L. donovani, comparing genome-wide protein synthesis patterns in the presence and absence of the HSP90-specific inhibitor radicicol and an ectopically expressed radicicol-resistant HSP90 variant. We find that ribosome-protected RNA faithfully maps open reading frames and represents 97% of the annotated protein-coding genes of L. donovani. Protein synthesis was found to correlate poorly with RNA steady-state levels, indicating a regulated translation as primary mechanism for HSP90-dependent gene expression. The results confirm inhibitory effects of HSP90 on the synthesis of Leishmania proteins that are associated with the pathogenic, intracellular stage of the parasite. Those include heat shock proteins, redox enzymes, virulence-enhancing surface proteins, proteolytic pathways, and a complete set of histones. Conversely, HSP90 promotes fatty acid synthesis enzymes. Complementing radicicol treatment with the radicicol-resistant HSP90rr variant revealed important off-target radicicol effects that control a large number of the above-listed proteins. Leishmania lacks gene-specific transcription regulation and relies on regulated translation instead. Our ribosome footprinting analysis demonstrates a controlling function of HSP90 in stage-specific protein synthesis but also significant, HSP90-independent effects of the inhibitor radicicol. IMPORTANCE Leishmania parasites cause severe illness in humans and animals. They exist in two developmental stages, insect form and mammalian form, which differ in shape and gene expression. By mapping and quantifying RNA fragments protected by protein synthesis complexes, we determined the rates of protein synthesis for >90% of all Leishmania proteins in response to the inhibition of a key regulatory protein, the 90-kDa heat shock protein. We find that Leishmania depends on a regulation of protein synthesis for controlling its gene expression and that heat shock protein 90 inhibition can trigger the developmental program from insect form to mammalian form of the pathogen.Eugenia BifeldStephan LorenzenKatharina BartschJuan-José VasquezT. Nicolai SiegelJoachim ClosAmerican Society for MicrobiologyarticleHSP90Leishmania donovaniradicicolribosome profilingMicrobiologyQR1-502ENmSystems, Vol 3, Iss 6 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	HSP90 Leishmania donovani radicicol ribosome profiling Microbiology QR1-502
spellingShingle	HSP90 Leishmania donovani radicicol ribosome profiling Microbiology QR1-502 Eugenia Bifeld Stephan Lorenzen Katharina Bartsch Juan-José Vasquez T. Nicolai Siegel Joachim Clos Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
description	ABSTRACT The 90-kDa heat shock protein (HSP90) of eukaryotes is a highly abundant and essential chaperone required for the maturation of regulatory and signal proteins. In the protozoan parasite Leishmania donovani, causative agent of the fatal visceral leishmaniasis, HSP90 activity is essential for cell proliferation and survival. Even more importantly, its inhibition causes life cycle progression from the insect stage to the pathogenic, mammalian stage. To unravel the molecular impact of HSP90 activity on the parasites’ gene expression, we performed a ribosome profiling analysis of L. donovani, comparing genome-wide protein synthesis patterns in the presence and absence of the HSP90-specific inhibitor radicicol and an ectopically expressed radicicol-resistant HSP90 variant. We find that ribosome-protected RNA faithfully maps open reading frames and represents 97% of the annotated protein-coding genes of L. donovani. Protein synthesis was found to correlate poorly with RNA steady-state levels, indicating a regulated translation as primary mechanism for HSP90-dependent gene expression. The results confirm inhibitory effects of HSP90 on the synthesis of Leishmania proteins that are associated with the pathogenic, intracellular stage of the parasite. Those include heat shock proteins, redox enzymes, virulence-enhancing surface proteins, proteolytic pathways, and a complete set of histones. Conversely, HSP90 promotes fatty acid synthesis enzymes. Complementing radicicol treatment with the radicicol-resistant HSP90rr variant revealed important off-target radicicol effects that control a large number of the above-listed proteins. Leishmania lacks gene-specific transcription regulation and relies on regulated translation instead. Our ribosome footprinting analysis demonstrates a controlling function of HSP90 in stage-specific protein synthesis but also significant, HSP90-independent effects of the inhibitor radicicol. IMPORTANCE Leishmania parasites cause severe illness in humans and animals. They exist in two developmental stages, insect form and mammalian form, which differ in shape and gene expression. By mapping and quantifying RNA fragments protected by protein synthesis complexes, we determined the rates of protein synthesis for >90% of all Leishmania proteins in response to the inhibition of a key regulatory protein, the 90-kDa heat shock protein. We find that Leishmania depends on a regulation of protein synthesis for controlling its gene expression and that heat shock protein 90 inhibition can trigger the developmental program from insect form to mammalian form of the pathogen.
format	article
author	Eugenia Bifeld Stephan Lorenzen Katharina Bartsch Juan-José Vasquez T. Nicolai Siegel Joachim Clos
author_facet	Eugenia Bifeld Stephan Lorenzen Katharina Bartsch Juan-José Vasquez T. Nicolai Siegel Joachim Clos
author_sort	Eugenia Bifeld
title	Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
title_short	Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
title_full	Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
title_fullStr	Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
title_full_unstemmed	Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>
title_sort	ribosome profiling reveals hsp90 inhibitor effects on stage-specific protein synthesis in <italic toggle="yes">leishmania donovani</italic>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/5dbeae5ae8294a028155e3a33ec51390
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Ribosome Profiling Reveals HSP90 Inhibitor Effects on Stage-Specific Protein Synthesis in <italic toggle="yes">Leishmania donovani</italic>

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