c trachomatis serovar l2  (ATCC)

Journal: PLOS One

Article Title: Screening a library of antibacterial compounds leads to discovery of novel inhibitors for Neisseria gonorrhoeae and Chlamydia trachomatis

doi: 10.1371/journal.pone.0340486

Figure Lengend Snippet: (A) Results of screening the antibacterial compounds library (containing 1,128 bioactive antibacterial agents), at a fixed concentration of 1 μM, against N. gonorrhoeae FA1090. Drugs exhibiting ≥ 90% inhibition of bacterial growth were considered positive hits (red). (B) Schematic of the screening process in this study, and the number of test agents at each step to finally obtain two molecules that are potent against N. gonorrhoeae and C. trachomatis .

Article Snippet: N. gonorrhoeae FA1090, C. trachomatis serovar L2 (ATCC VR-902B), and McCoy cell line were purchased from the American Type Culture Collection (ATCC).

Techniques: Concentration Assay, Inhibition

Journal: bioRxiv

Article Title: A lineage-specific heat-induced feedback loop controls HrcA to promote chlamydial fitness under stress

doi: 10.1101/2025.05.30.657042

Figure Lengend Snippet: (A) Under non-stressed conditions, HrcA binds to CIRCE elements and represses transcription from the KP1 and EP promoters. (B) Heat shock disrupts HrcA repression, allowing RNA polymerase (RNAP) to transcribe the hrcA-grpE-dnaK and groES-groEL operons. Arrows denote the extent of transcription. Note that dnaK is regulated by both the operon promoter and its own internal promoter (KP2). (C) Consensus and putative CIRCE sequences upstream of indicated heat shock genes in C. trachomatis (Ct) and Rhizobium meliloti (Rm) . Blue letters indicate nucleotide deviations from the consensus; lowercase letters denote mismatches between inverted repeats. The-35 core promoter elements for C. trachomatis genes are shown. (D) HrcA binds the ctl0271 promoter in chlamydiae. ChIP-qPCR was performed in biological triplicate using L2/NH-HrcA cultures treated with 10 nM ATC for 30 min at 37 °C. Anti-His antibody enrichment of the ctl0271 promoter was comparable to that of known HrcA targets ( groESL and dnaK P1), and significantly higher than enrichment at non-target promoters KP2 and E2P ( groEL2 promoter) , . (E) Expression of ctl0271 , dnaK , and groEL was reduced upon ATC-induced NH-HrcA overexpression in L2/NH-HrcA, as determined by qRT-PCR. (F) ctl0271 expression is induced in wild-type L2 cultured at 45 °C, as measured by qRT-PCR. (D–F) Data represent the mean ± SD of biological triplicates. (G, H) Based on results from panels B–F, the updated HrcA regulon includes ctl0271 (renamed hagF ) as its sixth target gene. (A, B, G, H) These panels were generated using a paid subscription to bioRender.

Article Snippet: Wild-type C. trachomatis serovar L2 strain 434/BU (L2) was obtained from the American Type Culture Collection (ATCC) and expanded in our laboratory using L929 cells as host cells.

Techniques: ChIP-qPCR, Expressing, Over Expression, Quantitative RT-PCR, Cell Culture, Generated

Journal: bioRxiv

Article Title: A lineage-specific heat-induced feedback loop controls HrcA to promote chlamydial fitness under stress

doi: 10.1101/2025.05.30.657042

Figure Lengend Snippet: (A) Effective silencing of hagF in C. trachomatis using deactivated CRISPR-associated protein 12 (dC12). L2/dC12-ntg expresses an ATC-inducible dC12 and a non-targeting guide RNA. L2/dC12-hagF-g1 and L2/dC12-hagF-g2 express dC12 and a guide RNA targeting hagF (g1 or g2). The left panel shows elevated dC12 transcript levels upon ATC induction in all three transformants. The right panel shows reduced hagF expression in ATC-treated L2/dC12-hagF-g1 and L2/dC12-hagF-g2 but not in L2/dC12-ntg. RNA levels were quantified by qRT-PCR. (B) ATC-induced dC12 expression has no effect on genome replication (left) or EB formation (right) in 37 °C cultures of L2/dC12-ntg. (C, D) ATC-induced hagF silencing has no detectable effect on genome replication (left) or EB formation (right) in 37 °C cultures of L2/dC12-hagF-g1 (C) and L2/dC12-hagF-g2 (D). (E) ATC-induced dC12 expression has no effect on genome replication (left) or EB formation (right) in 40.5 °C cultures of L2/dC12-ntg. (F, G) ATC-induced hagF silencing does not affect genome replication (left) but causes a substantial reduction in EB formation (right) in 40.5 °C cultures of L2/dC12-hagF-g1 (F) and L2/dC12-hagF-g2 (G). (B-G) ATC was added at 0 hpi. Genome copy number was quantified at the indicated times. EB yields were measured at 30 hpi for 37 °C cultures and at 40 hpi for 40.5 °C cultures.

Article Snippet: Wild-type C. trachomatis serovar L2 strain 434/BU (L2) was obtained from the American Type Culture Collection (ATCC) and expanded in our laboratory using L929 cells as host cells.

Techniques: CRISPR, Expressing, Quantitative RT-PCR

Journal: microPublication Biology

Article Title: UCK2-dependent conversion of cytidine to CTP is required for CTP uptake by Chlamydia trachomatis

doi: 10.17912/micropub.biology.001607

Figure Lengend Snippet: (A) Schematic view of the use of click chemistry to track nucleotide incorporation in nucleic acids. Cells are incubated with a nucleoside analog linked to an alkyne group (5-EC or N6pA). Nucleoside analogs are metabolized into nucleotides in the cytoplasm prior to incorporation into nucleic acids. After fixation and permeabilization, the click reaction between the alkyne and an azide group coupled to a fluorochrome, catalyzed by copper, allows to label the nucleotides. Created in https://BioRender.com. (B) HeLa cells treated with control (siCtrl) or siRNA targeting UCK2 (siUCK2) were infected with C. trachomatis (LGVL2 mCherry ) for 16 h before adding 5-EC or N6pA. Cells were fixed 6 h later, permeabilized and processed for covalent staining of the probes with Alexa-488 (green) and of DNA with DAPI (blue). Bacteria appear in red. The dotted white lines delimit the inclusions. Bar = 10 μm. (C-D) (Left) Fluorescence intensities of nucleoside analogs and of DAPI (C) or mCherry (D), in the nuclei and inclusions, respectively, were quantified in 50 cells per experiments. a.u. arbitrary units. P-values were determined with a paired t-test. (Right) Fluorescence from the nucleoside analogs normalized to the DAPI or to the mCherry signals, respectively. P-values were determined with a ratio paired t-test. Means ± SD of four independent experiments are shown. (E) HeLa cells treated with siCtrl or siUCK2 were infected with LGV L2 GFP (multiplicity of infection (MOI) ~ 0.3) and the percent of infected cells was determined 24 h later. A duplicate well was used to determine IFUs produced 48 hpi. Three independent experiments are represented. P-values were determined with a paired t-test (primary infection) and a ratio paired t-test (progeny).

Article Snippet: C. trachomatis serovar LGV L2 strain 434 (obtained from ATCC) stably expressing mCherry (click chemistry) or the green fluorescent protein (progeny) were used (Agaisse & Derré, 2013).

Techniques: Expressing, Incubation, Control, Infection, Staining, Bacteria, Fluorescence, Produced