GlcNAcylation confers protection against infection in through ubiquitination

• Published in: RSC advances 2018-06, Vol.8 (41), p.2389-231
• Main Authors: Vigneshwari, Loganathan, Balasubramaniam, Boopathi, Sethupathy, Sivasamy, Pandian, Shunmugiah Karutha, Balamurugan, Krishnaswamy
• Format: Article
• ISSN: 2046-2069
• DOI: 10.1039/c8ra00279g

Glycosylation is one of the most prevalent post-translational modifications in biological systems. In Caenorhabditis elegans , O -GlcNAcylation has been shown to be actively involved in the regulation of dauer formation and detoxification of toxins secreted by invading pathogens. On this backdrop, the present study is focused on understanding the role of O -GlcNAcylation in C. elegans during Staphylococcus aureus infection using a gel based proteomic approach. Results of time course killing assays with wild-type and mutants of glycosylation and comparison of results revealed an increase in the survival of the C. elegans oga-1 mutant when compared to wild-type N2 and the ogt-1 mutant. Increased survival of C. elegans N2 upon S. aureus infection in the presence of O -(2-acetamido-2-deoxy- d -glucopyranosylidenamino) N -phenylcarbamate (PUGNAc-an OGA inhibitor) further confirmed the involvement of O -GlcNAcylation in protecting C. elegans from infection. The two-dimensional gel-based proteomic analysis of the control and S. aureus infected C. elegans oga-1 mutant followed by mass spectrometric identification of differentially expressed proteins has been carried out. The results revealed that key proteins involved in ubiquitination such as Cullin-1 (CUL-1), Cullin-3 (CUL-3), BTB and MATH domain-containing protein 15 (BATH-15), ubiquitin-conjugating enzyme E2 variant 3 (UEV-3) and probable ubiquitin-conjugating enzyme E2 7 (UBC-7) are upregulated. Real-time PCR analysis further confirms the upregulation of genes encoding the above-mentioned proteins which are involved in the ubiquitin-mediated pathways in C. elegans . In addition, treatment of C. elegans wild-type N2 and the oga-1 mutant with PUGNAc + suramin and suramin (an ubiquitination inhibitor), respectively has resulted in increased sensitivity to S. aureus infection. Hence, it is presumed that upregulation of proteins involved in the ubiquitination pathway could be the key regulatory mechanism responsible for the enhanced survival of the oga-1 mutant during S. aureus infection. Two-dimensional gel-based proteomic approach unveiled that, O -GlcNAcylation protects Caenorhabditis elegans from Staphylococcus aureus infection by upregulating the proteins involved in ubiquitination pathway.