Article ; Online: Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress.
2017 Volume 7, Issue 1, Page(s) 1195
Abstract: ... shotgun proteomics, OxICAT and RNA-seq transcriptomics to analyse protein S-mycothiolation, reversible ... stress. In total, 58 S-mycothiolated proteins were identified under NaOCl stress that are involved ... detoxification. Protein S-mycothiolation was accompanied by MSH depletion in the thiol-metabolome. Quantification ...
| Abstract | 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. |
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| MeSH term(s) | Bacterial Proteins/metabolism ; Cysteine/metabolism ; Gene Expression Profiling ; Glycopeptides/metabolism ; Hypochlorous Acid/toxicity ; Inositol/metabolism ; Metabolic Networks and Pathways/genetics ; Metabolome ; Mycobacterium smegmatis/chemistry ; Mycobacterium smegmatis/drug effects ; Mycobacterium smegmatis/genetics ; Mycobacterium smegmatis/metabolism ; Oxidants/toxicity ; Oxidation-Reduction ; Protein Processing, Post-Translational ; Proteome/analysis ; Stress, Physiological |
| Chemical Substances | Bacterial Proteins ; Glycopeptides ; Oxidants ; Proteome ; mycothiol ; Inositol (4L6452S749) ; Hypochlorous Acid (712K4CDC10) ; Cysteine (K848JZ4886) |
| Language | English |
| Publishing date | 2017-04-26 |
| Publishing country | England |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 2615211-3 |
| ISSN | 2045-2322 ; 2045-2322 |
| ISSN (online) | 2045-2322 |
| ISSN | 2045-2322 |
| DOI | 10.1038/s41598-017-01179-4 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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