The chlamydial deubiquitinase Cdu1 supports recruitment of Golgi vesicles to the inclusion

Auer, Daniela; Hügelschäffer, Sophie Dorothea; Fischer, Annette; Rudel, Thomas

doi:10.1111/cmi.13136

Cited by 20 publications

(77 citation statements)

References 40 publications

(51 reference statements)

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“…Interestingly, we could often detect individual Chlamydia within close proximity to the inclusion membrane after feeding with NH 2 -ω-N 3 -C 6 -ceramides, possibly indicating an active docking to the inclusion membrane and an absorption of nutrition by C. trachomatis ( Supplementary Fig. 24 and Supplementary Movie 1) as has been hypothesized earlier 42 and reported in electron microscopy studies 40,43 . This behavior has previously been proposed as a mechanism by which RBs acquire nutrients including host lipids 44 and as an essential step in chlamydial development 42 .…”

Section: Imaging Interactions Of Bacteria and Intracellular Proteinssupporting

confidence: 75%

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

et al. 2020

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Expansion microscopy (ExM) enables super-resolution imaging of proteins and nucleic acids on conventional microscopes. However, imaging of details of the organization of lipid bilayers by light microscopy remains challenging. We introduce an unnatural short-chain azide- and amino-modified sphingolipid ceramide, which upon incorporation into membranes can be labeled by click chemistry and linked into hydrogels, followed by 4× to 10× expansion. Confocal and structured illumination microscopy (SIM) enable imaging of sphingolipids and their interactions with proteins in the plasma membrane and membrane of intracellular organelles with a spatial resolution of 10–20 nm. As our functionalized sphingolipids accumulate efficiently in pathogens, we use sphingolipid ExM to investigate bacterial infections of human HeLa229 cells by Neisseria gonorrhoeae, Chlamydia trachomatis and Simkania negevensis with a resolution so far only provided by electron microscopy. In particular, sphingolipid ExM allows us to visualize the inner and outer membrane of intracellular bacteria and determine their distance to 27.6 ± 7.7 nm.

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Section: Imaging Interactions Of Bacteria and Intracellular Proteinssupporting

confidence: 75%

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

et al. 2020

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“…Cdu1 is required for Golgi repositioning around the Ct inclusion (Pruneda et al, 2018; Auer et al, 2020). To understand how Cdu1 promotes Golgi redistribution, we first generated a cdu1 null strain in a Ct L2 (LGV L2 434 Bu) background by TargeTron mediated insertional mutagenesis (pDFTT3- aadA ) (Lowden et al, 2015) (S.…”

Section: Resultsmentioning

confidence: 99%

The acetylase activity of Cdu1 regulates bacterial exit from infected cells by protectingChlamydiaeffectors from degradation

Bastidas

Kędzior

Dolat

et al. 2023

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Many cellular processes are regulated by ubiquitin-mediated proteasomal degradation. Bacterial pathogens can regulate eukaryotic proteolysis through the delivery of proteins with de-ubiquitinating (DUB) activities. The obligate intracellular pathogenChlamydia trachomatissecretes Cdu1 (ChlaDUB1), a dual deubiquitinase and Lys-acetyltransferase, that promotes Golgi remodeling and survival of infected host cells presumably by regulating the ubiquitination of host and bacterial proteins. Here we determined that Cdu1's acetylase but not its DUB activity is important to protect Cdu1 from ubiquitin-mediated degradation. We further identified threeC. trachomatisproteins on the pathogen-containing vacuole (InaC, IpaM, and CTL0480) that required Cdu1's acetylase activity for protection from degradation and determined that Cdu1 and these Cdu1-protected proteins are required for optimal egress ofChlamydiafrom host cells. These findings highlight a non-canonical mechanism of pathogen-mediated protection of virulence factors from degradation after their delivery into host cells and the coordinated regulation of secreted effector proteins.

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“…Interestingly, we could often detect individual Chlamydia within close proximity to the inclusion membrane after feeding with NH2-ω-N3-C6-ceramides, possibly indicating an active docking to the inclusion membrane and an absorption of nutrition by C. trachomatis (Supplementary Figure S21 and Supplementary Movie 1) as has been hypothesized earlier 38 and reported in electron microscopy studies. 36,39 This behavior has previously been proposed as a mechanism by which RBs acquire nutrients including host lipids 40 and as an essential step in chlamydial development. 38 However, previous attempts to localize chlamydial particles in the inclusion required highly laborious techniques such as Serial block-face scanning electron microscopy.…”

Section: Imaging Interactions Of Bacteria and Intracellular Proteinsmentioning

confidence: 87%

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

Goetz

Kunz

Fink

et al. 2020

Preprint

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Expansion microscopy (ExM) enables super-resolution imaging of proteins and nucleic acids on conventional microscopes. However, imaging of details of the organization of lipid bilayers by light microscopy remains challenging. We introduce an azide-and amino-modified sphingolipid ceramide, which upon incorporation into membranes can be labeled by click chemistry and linked into hydrogels, followed by 4x to 10x expansion. Confocal and structured illumination microscopy (SIM) enabled imaging of sphingolipids and their interactions with proteins in the membrane of intracellular organelles with a spatial resolution of 10-20 nm. Because sphingolipids accumulated efficiently in pathogens we used sphingolipid ExM to investigate bacterial infections of human HeLa229 cells by Neisseria gonorrhoeae, Chlamydia trachomatis and Simkania negevensis with a resolution so far only provided by electron microscopy. In particular, sphingolipid ExM allowed us to visualize the inner and outer membrane of intracellular bacteria and determine their distance to 27.6 ± 7.7 nm. The Supporting Information is available free of charge on the ACS Publications website. Supplementary Figures S1-S23. Supplementary Movie 1: 10x ExM SIM z-stack of Hela229 cells infected with Chlamydia trachomatis for 24 h, fed with α-NH2-ω-N3-C6-ceramide, fixed, permeabilized and stained with DBCO-Alexa Fluor 488. Chlamydia are clearly located at the inclusion membrane. Scale bar, 10 µm.

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The chlamydial deubiquitinase Cdu1 supports recruitment of Golgi vesicles to the inclusion

Cited by 20 publications

References 40 publications

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

The acetylase activity of Cdu1 regulates bacterial exit from infected cells by protectingChlamydiaeffectors from degradation

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

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