Cargando…

Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress

Mycothiol (MSH) is the major low molecular weight (LMW) thiol in Actinomycetes. Here, we used shotgun proteomics, OxICAT and RNA-seq transcriptomics to analyse protein S-mycothiolation, reversible thiol-oxidations and their impact on gene expression in Mycobacterium smegmatis under hypochlorite stre...

Descripción completa

Detalles Bibliográficos
Autores principales: Hillion, Melanie, Bernhardt, Jörg, Busche, Tobias, Rossius, Martina, Maaß, Sandra, Becher, Dörte, Rawat, Mamta, Wirtz, Markus, Hell, Rüdiger, Rückert, Christian, Kalinowski, Jörn, Antelmann, Haike
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5430705/
https://www.ncbi.nlm.nih.gov/pubmed/28446771
http://dx.doi.org/10.1038/s41598-017-01179-4
_version_ 1783236275597213696
author Hillion, Melanie
Bernhardt, Jörg
Busche, Tobias
Rossius, Martina
Maaß, Sandra
Becher, Dörte
Rawat, Mamta
Wirtz, Markus
Hell, Rüdiger
Rückert, Christian
Kalinowski, Jörn
Antelmann, Haike
author_facet Hillion, Melanie
Bernhardt, Jörg
Busche, Tobias
Rossius, Martina
Maaß, Sandra
Becher, Dörte
Rawat, Mamta
Wirtz, Markus
Hell, Rüdiger
Rückert, Christian
Kalinowski, Jörn
Antelmann, Haike
author_sort Hillion, Melanie
collection PubMed
description Mycothiol (MSH) is the major low molecular weight (LMW) thiol in Actinomycetes. Here, we used shotgun proteomics, OxICAT and RNA-seq transcriptomics to analyse protein S-mycothiolation, reversible thiol-oxidations and their impact on gene expression in Mycobacterium smegmatis under hypochlorite stress. In total, 58 S-mycothiolated proteins were identified under NaOCl stress that are involved in energy metabolism, fatty acid and mycolic acid biosynthesis, protein translation, redox regulation and detoxification. Protein S-mycothiolation was accompanied by MSH depletion in the thiol-metabolome. Quantification of the redox state of 1098 Cys residues using OxICAT revealed that 381 Cys residues (33.6%) showed >10% increased oxidations under NaOCl stress, which overlapped with 40 S-mycothiolated Cys-peptides. The absence of MSH resulted in a higher basal oxidation level of 338 Cys residues (41.1%). The RseA and RshA anti-sigma factors and the Zur and NrdR repressors were identified as NaOCl-sensitive proteins and their oxidation resulted in an up-regulation of the SigH, SigE, Zur and NrdR regulons in the RNA-seq transcriptome. In conclusion, we show here that NaOCl stress causes widespread thiol-oxidation including protein S-mycothiolation resulting in induction of antioxidant defense mechanisms in M. smegmatis. Our results further reveal that MSH is important to maintain the reduced state of protein thiols.
format Online
Article
Text
id pubmed-5430705
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-54307052017-05-16 Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress Hillion, Melanie Bernhardt, Jörg Busche, Tobias Rossius, Martina Maaß, Sandra Becher, Dörte Rawat, Mamta Wirtz, Markus Hell, Rüdiger Rückert, Christian Kalinowski, Jörn Antelmann, Haike Sci Rep Article Mycothiol (MSH) is the major low molecular weight (LMW) thiol in Actinomycetes. Here, we used shotgun proteomics, OxICAT and RNA-seq transcriptomics to analyse protein S-mycothiolation, reversible thiol-oxidations and their impact on gene expression in Mycobacterium smegmatis under hypochlorite stress. In total, 58 S-mycothiolated proteins were identified under NaOCl stress that are involved in energy metabolism, fatty acid and mycolic acid biosynthesis, protein translation, redox regulation and detoxification. Protein S-mycothiolation was accompanied by MSH depletion in the thiol-metabolome. Quantification of the redox state of 1098 Cys residues using OxICAT revealed that 381 Cys residues (33.6%) showed >10% increased oxidations under NaOCl stress, which overlapped with 40 S-mycothiolated Cys-peptides. The absence of MSH resulted in a higher basal oxidation level of 338 Cys residues (41.1%). The RseA and RshA anti-sigma factors and the Zur and NrdR repressors were identified as NaOCl-sensitive proteins and their oxidation resulted in an up-regulation of the SigH, SigE, Zur and NrdR regulons in the RNA-seq transcriptome. In conclusion, we show here that NaOCl stress causes widespread thiol-oxidation including protein S-mycothiolation resulting in induction of antioxidant defense mechanisms in M. smegmatis. Our results further reveal that MSH is important to maintain the reduced state of protein thiols. Nature Publishing Group UK 2017-04-26 /pmc/articles/PMC5430705/ /pubmed/28446771 http://dx.doi.org/10.1038/s41598-017-01179-4 Text en © The Author(s) 2017 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
Hillion, Melanie
Bernhardt, Jörg
Busche, Tobias
Rossius, Martina
Maaß, Sandra
Becher, Dörte
Rawat, Mamta
Wirtz, Markus
Hell, Rüdiger
Rückert, Christian
Kalinowski, Jörn
Antelmann, Haike
Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title_full Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title_fullStr Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title_full_unstemmed Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title_short Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
title_sort monitoring global protein thiol-oxidation and protein s-mycothiolation in mycobacterium smegmatis under hypochlorite stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5430705/
https://www.ncbi.nlm.nih.gov/pubmed/28446771
http://dx.doi.org/10.1038/s41598-017-01179-4
work_keys_str_mv AT hillionmelanie monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT bernhardtjorg monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT buschetobias monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT rossiusmartina monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT maaßsandra monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT becherdorte monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT rawatmamta monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT wirtzmarkus monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT hellrudiger monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT ruckertchristian monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT kalinowskijorn monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress
AT antelmannhaike monitoringglobalproteinthioloxidationandproteinsmycothiolationinmycobacteriumsmegmatisunderhypochloritestress