Monitoring global protein thiol-oxidation and protein S-mycothiolation in Mycobacterium smegmatis under hypochlorite stress
Name
Hillion-2017-Monitoring global protein thiol-o.pdf
Size
8.1 MB
Format
Adobe PDF
Checksum (MD5)
4d76863c70c890b5da405b7596d262ef
Author(s)
Hillion, Melanie
Busche, Tobias
Rossius, Martina
Rawat, Mamta
Wirtz, Markus
Antelmann, Haike
Bernhardt, Jorg
Maas, Sandra
Becher, Dorte
Hell, Ruediger
Date Issued
April 2017
Journal
Scientific Reports
Publisher
Nature Publishing Group
Citation
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
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution 4.0 International License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/s41598-017-01179-4
