biostudies-literatureLewis JMNational Health and Medical Research CouncilAustralian Research Council1762-1778https://www.ebi.ac.uk/biostudies/studies/S-EPMC10243306biostudies-literatureUnknown22(6)The process of O-linked protein glycosylation is highly conserved across the <i>Burkholderia</i> genus and mediated by the oligosaccharyltransferase PglL. While our understanding of <i>Burkholderia</i> glycoproteomes has increased in recent years, little is known about how <i>Burkholderia</i> species respond to modulations in glycosylation. Utilizing CRISPR interference (CRISPRi), we explored the impact of silencing of O-linked glycosylation across four species of <i>Burkholderia</i>; <i>Burkholderia cenocepacia</i> K56-2, <i>Burkholderia diffusa</i> MSMB375, <i>Burkholderia multivorans</i> ATCC17616, and <i>Burkholderia thailandensis</i> E264. Proteomic and glycoproteomic analyses revealed that while CRISPRi enabled inducible silencing of PglL, this did not abolish glycosylation, nor recaJournal of proteome researchCRISPRi-Mediated Silencing of <i>Burkholderia</i> O-Linked Glycosylation Systems Enables the Depletion of Glycosylation Yet Results in Modest Proteome Impacts.PMC102433062018980FT200100270DP210100362Lewis JMScott NEfalseCRISPRi-Mediated Silencing of <i>Burkholderia</i> O-Linked Glycosylation Systems Enables the Depletion of Glycosylation Yet Results in Modest Proteome Impacts.The process of O-linked protein glycosylation is highly conserved across the <i>Burkholderia</i> genus and mediated by the oligosaccharyltransferase PglL. While our understanding of <i>Burkholderia</i> glycoproteomes has increased in recent years, little is known about how <i>Burkholderia</i> species respond to modulations in glycosylation. Utilizing CRISPR interference (CRISPRi), we explored the impact of silencing of O-linked glycosylation across four species of <i>Burkholderia</i>; <i>Burkholderia cenocepacia</i> K56-2, <i>Burkholderia diffusa</i> MSMB375, <i>Burkholderia multivorans</i> ATCC17616, and <i>Burkholderia thailandensis</i> E264. Proteomic and glycoproteomic analyses revealed that while CRISPRi enabled inducible silencing of PglL, this did not abolish glycosylation, nor reca2023-01-01T00:00:00Z2023 Jun2026-06-01T21:47:14.414Z2025-04-04T19:12:16.996ZS-EPMC102433063699511410.1021/acs.jproteome.2c00790