Time course infection of HeLa cells by Chlamydia tracomatis
Ontology highlight
ABSTRACT:
INSTRUMENT(S):
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Permanent Cell Line Cell
SUBMITTER: 
Allan Stensballe
LAB HEAD: Allan Stensballe
PROVIDER: PXD001457 | Pride | 2018-01-08
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| Pride_MaxQ_fulltrypsin.zip | Other | |||
| Pride_MaxQ_semitrypsin.zip | Other | |||
| SB36_01_hela0hrs.raw | Raw | |||
| SB36_02_hela0hrs.raw | Raw | |||
| SB36_03_hela0hrs.raw | Raw |
Items per page: 5 1 - 5 of 23 |
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Project description:Lysosomal acid lipase (LAL) is the key enzyme of lysosomal lipid hydrolysis, which degrades cholesteryl esters (CE), triacylglycerols (TG), diacylglycerols (DG), and retinyl esters. The role of LAL in various cellular processes has mostly been studied in LAL-deficient (Lal-/-) mice, which share phenotypical characteristics with humans suffering from LAL deficiency. In vitro, the cell-specific functions of LAL have been commonly investigated by using the LAL inhibitors Lalistat-1 (L1) and Lalistat-2 (L2). Here, we show that pharmacological LAL inhibition but not genetic loss of LAL impairs isoproterenol-stimulated lipolysis and neutral TG hydrolase (TGH) and CE hydrolase (CEH) activities in mature adipocytes, indicating that L1 and L2 inhibit other lipid hydrolases apart from LAL. Since adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) are the major enzymes that degrade cytosolic TG and CE, respectively, at neutral pH, we hypothesized that L1 and L2 also inhibit ATGL and/or HSL through off-target effects. In fact, both inhibitors drastically reduced neutral CEH activity in cells overexpressing mouse and human HSL and neutral TGH activity in cells overexpressing mouse and human ATGL, albeit to a lesser extent. By performing serine hydrolase-specific activity-based labeling in combination with quantitative proteomics, we confirmed that L2 inhibits HSL and other lipid hydrolases, whereas L1 treatment results in less pronounced inhibition of neutral lipid hydrolases. These results demonstrate that commonly used concentrations of L2 (and L1) are not suitable for investigating the role of LAL-specific lipolysis in lysosomal function, signaling pathways, and autophagy.
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2009-01-15 | GSE10199 | GEO
Project description:Chlamydia trachomatis is the leading cause of bacterial sexually transmitted infections in the US and preventable blindness worldwide. This obligate intracellular pathogen replicates within a membrane-bound inclusion, but how it acquires nutrients from the host while avoiding detection by the innate immune system is incompletely understood. Chlamydia accomplishes this in part through the translocation of a unique set of effectors into the inclusion membrane, the inclusion membrane proteins (Incs). Incs are ideally poised at the host-pathogen interface to reprogram host signaling by redirecting proteins or organelles to the inclusion. Using a combination of co-affinity purification, immunofluorescence confocal imaging, and proteomics, we characterize the interaction between an early-expressed Inc of unknown function, Tri1, and tumor necrosis factor receptor associated factor 7 (TRAF7). TRAF7 is a multi-domain protein with a RING finger ubiquitin ligase domain and a C-terminal WD40 domain, the latter is mutated in a subset of tumors. TRAF7 regulates several innate immune signaling pathways associated with C. trachomatis infection. We demonstrate that Tri1 and TRAF7 specifically interact during infection and that TRAF7 is recruited to the inclusion. We further show that the predicted coiled-coil domain Tri1 is necessary and sufficient to interact with the TRAF7 WD40 domain. Finally, we demonstrate that Tri1 displaces native TRAF7 binding partners, MEKK2 and MEKK3. Together, our results suggest that by displacing the TRAF7 native binding partners MEKK2 and/or MEKK3, Tri1 has the capacity to alter TRAF7 signaling during infection.
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2010-09-16 | E-GEOD-24119 | biostudies-arrayexpress
Project description:C. trachomatis possess a cryptic 7.5 kb plasmid of unknown function. Here we describe a comprehensive molecular and biological characterization of the naturally occurring plasmidless human Chlamydia trachomatis strain L2 (25667R). We found that despite minimal chromosomal polymorphisms the LGV L2 (25667R) strain was indistinguishable from the L2 (434) plasmid positive strain in its in vitro infectivity characteristics such as growth kinetics, plaquing efficiency, and plaque size. The primary in vitro phenotypic differences between L2 (434) and L2 (25667R) were the accumulation of glycogen granules in the inclusion matrix and the lack of the typical intra-inclusion Brownian-like movement characteristic of C. trachomatis strains. Conversely, we observed a marked difference between the two strains in their ability to colonize and infect the female mouse genital tract. The ID50 of the L2 (25667R) plasmidless strain was 500 fold greater (1.XX x 10X IFU) than the L2 (434) plasmid bearing strain (1. XX x 10X IFU). Transcriptome analysis of the two strains clearly demonstrated a decrease in transcript levels of a subset of chromosomal genes for the L2 (25667R) strain. Among those genes was glgA which encodes for glycogen synthase; a finding consistent with the failure of the L2 (25667R) strain to accumulate glycogen granules. Collectively, these findings support an important role for the plasmid in in vivo infectivity and suggest that this virulence characteristic might be controlled by the plasmids ability to regulate the expression of specific chromosomal genes. These results also support an important role for the plasmid in the pathogenesis of human infection and disease. Keywords: strain comparison C. trachomatis strain L2-25667R compared to strain L2-434
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