Integrated Succinylome and Metabolome Profiling Reveals Crucial Role of S-Ribosylhomocysteine Lyase in Quorum Sensing and Metabolism of Aeromonas hydrophila*

Yao, Zujie; Guo, Zhuang; Wang, Yuqian; Li, Wanxin; Fu, Yuying; Lin, Yongjun; Lin, Wenxiong; Lin, Xiangmin

doi:10.1074/mcp.ra118.001035

Cited by 44 publications

(56 citation statements)

References 59 publications

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“…We previously reported that A0KHS0 (AHA_1280) and A0KHS1 (AHA_1281) have several lysine succinylation modifications sites but not A0KHR9 (AHA_1279). Whether this post-translational modification could affect the size of the pore of these proteins and ultimately influence their biological functions need to be further investigated (Yao et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

The characteristics of antibiotic resistance and phenotypes in 29 outer‐membrane protein mutant strains in Aeromonas hydrophila

Wang

et al. 2019

Environmental Microbiology

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Summary Although many typical outer‐membrane proteins (OMPs) have been well characterized, the biological functions of many OMPs remain largely elusive. In this study, we successfully constructed 29 OMP knockout strains in the pathogen Aeromonas hydrophila, which account for about 50% of all predicted OMPs in this bacterial species. We then further validated the antibiotics' susceptibility characteristics against 20 antimicrobial reagents in these mutants considering several phenotypes. Our results showed that a total of 22 OMP mutants affected the susceptibility to at least one antibiotic. The deletion of some OMPs, such as ΔlamB and ΔbamA, revealed very important roles in the resistance to certain antibiotics. However, not a single OMP mutant presented a constant behaviour to all of the tested antibiotics, suggesting the existence of a complex intercellular regulation mechanism and a protein–protein interaction network underlying the OMP homeostasis in the presence of antibiotics. Meanwhile, some OMP mutants also affected biofilm formation, ECPase and haemolytic activity, and carbon resources utilization. This report demonstrates the biological functions of OMPs on a large scale and most of results have not been reported in A. hydrophila.

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Section: Discussionmentioning

confidence: 99%

The characteristics of antibiotic resistance and phenotypes in 29 outer‐membrane protein mutant strains in Aeromonas hydrophila

Wang

et al. 2019

Environmental Microbiology

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“…Expression of NqrF was detected as previously described (Yao et al ., ). Briefly, the whole bacterial lysates were separated by 10% SDS‐PAGE and the proteins in the gel were transferred onto nitrocellulose (NC) membranes by electroblotting at a constant voltage of 80 V for 2 h. The membranes were blocked overnight with 5% nonfat milk in Tris‐buffered saline buffer containing 0.05% Tween‐20 (TBST) at 4 °C.…”

Section: Methodsmentioning

confidence: 97%

“…Expression of NqrF was detected as previously described (Yao et al, 2019). Briefly, the whole bacterial lysates were separated by 10% SDS-PAGE and the proteins in the gel were transferred onto nitrocellulose (NC) membranes by electroblotting at a constant voltage of 80 V for 2 h. The membranes were blocked overnight with 5% nonfat milk in Tris-buffered saline buffer containing 0.05% Tween-20 (TBST) at 4 C. Then, the membranes were incubated with 1:50 of the primary mouse anti-NqrF after rinsing three times for 10 min with TBST buffer and subsequently incubated with a horseradish peroxidase-linked secondary antibody.…”

Section: Western Blot Analysismentioning

confidence: 99%

Reduced redox‐dependent mechanism and glucose‐mediated reversal in gentamicin‐resistant Vibrio alginolyticus

Zhang

Wang

Jiang

et al. 2019

Environmental Microbiology

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Summary Strategy of managing antibiotic‐resistant Vibrio alginolyticus, a bacterial pathogen that threatens human health and animal farming, is not available due to the lack of knowledge about the underlying mechanism of antibiotic resistance. Here, we showed that gentamicin‐resistant V. alginolyticus (VA‐RGEN) has four mutations on metabolism and one mutation on a two‐component system by whole‐genome and PCR‐based sequencing, indicating the metabolic shift in VA‐RGEN. Thus, metabolic profile was investigated by GC–MS based metabolomics. Glucose was identified as a crucial biomarker, whose abundance was decreased in VA‐RGEN. Further analysis with iPath, and gene expression and enzyme activity of the pyruvate cycle (the P cycle) demonstrated a global depressed metabolic pathway network in VA‐RGEN. Consistently, NADH, sodium‐pumping NADH:ubiquinone oxidoreductase (Na(+)‐NQR) system, membrane potential and intracellular gentamicin were decreased in VA‐RGEN. These findings indicate that the reduced redox state contributes to antibiotic resistance. Interestingly, exogenous glucose potentiated gentamicin to efficiently kill VA‐RGEN through the promotion of the P cycle, NADH, membrane potential and intracellular gentamicin. The potentiation was further confirmed in a zebrafish model. These results indicate that the gentamicin resistance reduces the P cycle and Na(+)‐NQR system and thereby decreases redox state, membrane potential and gentamicin uptake, which can be reversed by exogenous glucose.

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“…The dynamic reversible modification of target proteins may affect the PPI, thus contributing to the regulation of metabolic processes [35]. The modified proteins were then evaluated for PPI networks and clustered into KEGG pathways using STRING (minimum required interaction score is 0.7).…”

Section: Plos Pathogensmentioning

confidence: 99%

S-glutathionylation proteome profiling reveals a crucial role of a thioredoxin-like protein in interspecies competition and cariogenecity of Streptococcus mutans

Zhang

Cheng

et al. 2020

PLoS Pathog

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S-glutathionylation is an important post-translational modification (PTM) process that targets protein cysteine thiols by the addition of glutathione (GSH). This modification can prevent proteolysis caused by the excessive oxidation of protein cysteine residues under oxidative or nitrosative stress conditions. Recent studies have suggested that protein S-glutathionylation plays an essential role in the control of cell-signaling pathways by affecting the protein function in bacteria and even humans. In this study, we investigated the effects of Sglutathionylation on physiological regulation within Streptococcus mutans, the primary etiological agent of human dental caries. To determine the S-glutathionylated proteins in bacteria, the Cys reactive isobaric reagent iodoacetyl Tandem Mass Tag (iodoTMT) was used to label the S-glutathionylated Cys site, and an anti-TMT antibody-conjugated resin was used to enrich the modified peptides. Proteome profiling identified a total of 357 glutathionylated cysteine residues on 239 proteins. Functional enrichment analysis indicated that these Sglutathionylated proteins were involved in diverse important biological processes, such as pyruvate metabolism and glycolysis. Furthermore, we studied a thioredoxin-like protein (Tlp) to explore the effect of S-glutathionylation on interspecies competition between oral streptococcal biofilms. Through site mutagenesis, it was proved that glutathionylation on Cys41 residue of Tlp is crucial to protect S. mutans from oxidative stress and compete with S. sanguinis and S. gordonii. An addition rat caries model showed that the loss of S-glutathionylation attenuated the cariogenicity of S. mutans. Taken together, our study provides an insight into the S-glutathionylation of bacterial proteins and the regulation of oxidative stress resistance and interspecies competition.

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Integrated Succinylome and Metabolome Profiling Reveals Crucial Role of S-Ribosylhomocysteine Lyase in Quorum Sensing and Metabolism of Aeromonas hydrophila*

Cited by 44 publications

References 59 publications

The characteristics of antibiotic resistance and phenotypes in 29 outer‐membrane protein mutant strains in Aeromonas hydrophila

The characteristics of antibiotic resistance and phenotypes in 29 outer‐membrane protein mutant strains in Aeromonas hydrophila

Reduced redox‐dependent mechanism and glucose‐mediated reversal in gentamicin‐resistant Vibrio alginolyticus

S-glutathionylation proteome profiling reveals a crucial role of a thioredoxin-like protein in interspecies competition and cariogenecity of Streptococcus mutans

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