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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2017
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| Online Access: | http://hdl.handle.net/1721.1/110027 |
| _version_ | 1811085852768468992 |
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| author | Hillion, Melanie Busche, Tobias Rossius, Martina Rawat, Mamta Wirtz, Markus Antelmann, Haike Bernhardt, Jorg Maas, Sandra Becher, Dorte Hell, Ruediger Kalinowski, Jorn Rueckert, Christian |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Hillion, Melanie Busche, Tobias Rossius, Martina Rawat, Mamta Wirtz, Markus Antelmann, Haike Bernhardt, Jorg Maas, Sandra Becher, Dorte Hell, Ruediger Kalinowski, Jorn Rueckert, Christian |
| author_sort | Hillion, Melanie |
| collection | MIT |
| 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. |
| first_indexed | 2024-09-23T13:16:34Z |
| format | Article |
| id | mit-1721.1/110027 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:16:34Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1100272022-09-28T13:06:10Z Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress Hillion, Melanie Busche, Tobias Rossius, Martina Rawat, Mamta Wirtz, Markus Antelmann, Haike Bernhardt, Jorg Maas, Sandra Becher, Dorte Hell, Ruediger Kalinowski, Jorn Rueckert, Christian Massachusetts Institute of Technology. Department of Biology Rueckert, Christian 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. 2017-06-19T20:07:13Z 2017-06-19T20:07:13Z 2017-04 2016-12 Article http://purl.org/eprint/type/JournalArticle 2045-2322 http://hdl.handle.net/1721.1/110027 Hillion, Melanie; Bernhardt, Jörg; Busche, Tobias; Rossius, Martina; Maaß, Sandra; Becher, Dörte and Rawat, Mamta et al. “Monitoring Global Protein Thiol-Oxidation and Protein S-Mycothiolation in Mycobacterium Smegmatis Under Hypochlorite Stress.” Scientific Reports 7, no. 1 (April 2017): 1195 © 2017 Macmillan Publishers Limited, part of Springer Nature en_US http://dx.doi.org/10.1038/s41598-017-01179-4 Scientific Reports Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group Nature |
| spellingShingle | Hillion, Melanie Busche, Tobias Rossius, Martina Rawat, Mamta Wirtz, Markus Antelmann, Haike Bernhardt, Jorg Maas, Sandra Becher, Dorte Hell, Ruediger Kalinowski, Jorn Rueckert, Christian 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 |
| url | http://hdl.handle.net/1721.1/110027 |
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