Proximity Labeling To Map Host-Pathogen Interactions at the Membrane of a Bacterium-Containing Vacuole in Chlamydia trachomatis-Infected Human Cells

Olson, Macy G.; Widner, Ray E.; Jorgenson, Lisa M; Lawrence, Alyssa; Lagundžin, Dragana; Woods, Nicholas T.; Ouellette, Scot P.; Rucks, Elizabeth A.

doi:10.1128/iai.00537-19

Cited by 30 publications

(83 citation statements)

References 94 publications

(222 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our observation that silencing GFPT expression increases bacterial diameter strongly supports the hypothesis that UDP‐GlcNAc, or an intermediate along the HBP, is hijacked into the inclusion to fuel bacterial division, possibly by feeding peptidoglycan synthesis. Interestingly, a proximity‐based labeling assay recently described an enrichment of TG2 and GFPT1 around the inclusion, suggesting that TG2 activation and GFPT modification might be most efficient in proximity of the bacteria‐containing compartment (Olson et al , ). One recent publication showed that UDP‐GlcNAc is also used during infection to post‐translationally modify the intermediate filament vimentin and this also could contribute to significant UDP‐GlcNAc consumption in C. trachomatis infection (Tarbet et al , ).…”

Section: Discussionmentioning

confidence: 99%

Infection‐driven activation of transglutaminase 2 boosts glucose uptake and hexosamine biosynthesis in epithelial cells

Maffei

Laverrière

et al. 2020

The EMBO Journal

View full text Add to dashboard Cite

Transglutaminase 2 (TG2) is a ubiquitously expressed enzyme with transamidating activity. We report here that both expression and activity of TG2 are enhanced in mammalian epithelial cells infected with the obligate intracellular bacteria Chlamydia trachomatis. Genetic or pharmacological inhibition of TG2 impairs bacterial development. We show that TG2 increases glucose import by upregulating the transcription of the glucose transporter genes GLUT-1 and GLUT-3. Furthermore, TG2 activation drives one specific glucose-dependent pathway in the host, i.e., hexosamine biosynthesis. Mechanistically, we identify the glucosamine:fructose-6phosphate amidotransferase (GFPT) among the substrates of TG2. GFPT modification by TG2 increases its enzymatic activity, resulting in higher levels of UDP-N-acetylglucosamine biosynthesis and protein O-GlcNAcylation. The correlation between TG2 transamidating activity and O-GlcNAcylation is disrupted in infected cells because host hexosamine biosynthesis is being exploited by the bacteria, in particular to assist their division. In conclusion, our work establishes TG2 as a key player in controlling glucose-derived metabolic pathways in mammalian cells, themselves hijacked by C. trachomatis to sustain their own metabolic needs.

show abstract

Section: Discussionmentioning

confidence: 99%

Infection‐driven activation of transglutaminase 2 boosts glucose uptake and hexosamine biosynthesis in epithelial cells

Maffei

Laverrière

et al. 2020

The EMBO Journal

View full text Add to dashboard Cite

show abstract

“…An alternative proximity labeling (PL) approach, relies of the translational fusion of engineered APEX to biotinylate proteins in the vicinity of the bait [85]. In the presence of H 2 O 2 , APEX catalyzes the oxidation of biotin-phenol to the highly reactive and short-lived biotin-phenoxyl radical that subsequently reacts with electron-rich amino acids (>98% tyrosine and <2% tryptophan, cysteine [86]) of proximal proteins in a range of~20 nm [86], resulting in their biotinylation.…”

Section: Proximity-dependent Labeling Approachesmentioning

confidence: 99%

Keeping in Touch with Type-III Secretion System Effectors: Mass Spectrometry-Based Proteomics to Study Effector–Host Protein–Protein Interactions

Meyer

Ryck

Goormachtig

et al. 2020

IJMS

View full text Add to dashboard Cite

Manipulation of host cellular processes by translocated bacterial effectors is key to the success of bacterial pathogens and some symbionts. Therefore, a comprehensive understanding of effectors is of critical importance to understand infection biology. It has become increasingly clear that the identification of host protein targets contributes invaluable knowledge to the characterization of effector function during pathogenesis. Recent advances in mapping protein–protein interaction networks by means of mass spectrometry-based interactomics have enabled the identification of host targets at large-scale. In this review, we highlight mass spectrometry-driven proteomics strategies and recent advances to elucidate type-III secretion system effector–host protein–protein interactions. Furthermore, we highlight approaches for defining spatial and temporal effector–host interactions, and discuss possible avenues for studying natively delivered effectors in the context of infection. Overall, the knowledge gained when unravelling effector complexation with host factors will provide novel opportunities to control infectious disease outcomes.

show abstract

“…Lately, the Ting group further enhanced the labeling efficiency of BioID via directed evolution of the biotin ligase (now named TurboID) [ 56 ]. Thus far, proximity-labeling methods have contributed the discovery of several novel protein-protein interactions at the host-pathogen interface [ 57 , 58 , 59 ]. Last but not least, chemical crosslinking in principle can stabilize protein complexes by introducing covalent interactions.…”

Section: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Contributions of Mass Spectrometry-Based Proteomics to Understanding Salmonella-Host Interactions

et al. 2020

View full text Add to dashboard Cite

As a model pathogen, Salmonella invades both phagocytic and non-phagocytic host cells and adopts an intracellular lifestyle in a membrane-bound compartment during infection. Therefore, a systemic overview of Salmonella adaptations to distinct host cells together with host remodeling will assist us in charting the landscape of host-pathogen interactions. Central to the Salmonella-host interplay are bacterial virulence factors (effectors) that are injected into host cells by type III secretion systems (T3SSs). Despite great progress, functional studies of bacterial effectors have experienced daunting challenges as well. In the last decade, mass spectrometry-based proteomics has evolved into a powerful technological platform that can quantitatively measure thousands of proteins in terms of their expression as well as post-translational modifications. Here, we will review the applications of high-throughput proteomic technologies in understanding the dynamic reprogramming of both Salmonella and host proteomes during the course of infection. Furthermore, we will summarize the progress in utilizing affinity purification-mass spectrometry to screen for host substrates of Salmonella T3SS effectors. Finally, we will critically discuss some limitations/challenges with current proteomic platforms in the context of host-pathogen interactions and highlight some emerging technologies that may offer the promise of tackling these problems.

show abstract

Proximity Labeling To Map Host-Pathogen Interactions at the Membrane of a Bacterium-Containing Vacuole in Chlamydia trachomatis-Infected Human Cells

Cited by 30 publications

References 94 publications

Infection‐driven activation of transglutaminase 2 boosts glucose uptake and hexosamine biosynthesis in epithelial cells

Infection‐driven activation of transglutaminase 2 boosts glucose uptake and hexosamine biosynthesis in epithelial cells

Keeping in Touch with Type-III Secretion System Effectors: Mass Spectrometry-Based Proteomics to Study Effector–Host Protein–Protein Interactions

Contributions of Mass Spectrometry-Based Proteomics to Understanding Salmonella-Host Interactions

Contact Info

Product

Resources

About