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atcc 27853 wt strain  (ATCC)


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    ATCC atcc 27853 wt strain
    Temperature downshifts enhanced biofilm formation in the majority of Pseudomonas aeruginosa clinical and environmental isolates. A , biofilm biomass of 52 clinical P. aeruginosa isolates, P. aeruginosa PAO1, and four environmental isolates following culturing statically at RT and 37 °C. M1: ATCC 27853. M35: PA150663 . M54: PA152541 . B , dot plot summarizing the biofilm patterns of P. aeruginosa isolates at RT and 37 °C. Black : environmental isolates. The two dash lines represent the threshold of 2-fold. Blue : strains formed two-fold or more biofilm at RT relative to 37 °C. Red : strains formed two-fold or more biofilm at 37 °C relative to RT. See related for bar graph with statistical analysis. C , biofilm biomass of ATCC 27853, PAO1, PA150663 , and PA152541 at a temperature gradient from 15 °C to 42 °C at 48 h. D , biofilm formation of P. aeruginosa <t>ATCC</t> <t>27853</t> and PAO1 at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).
    Atcc 27853 Wt Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 22879 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    atcc 27853 wt strain - by Bioz Stars, 2026-10
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    1) Product Images from "Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module"

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2025.111086

    Temperature downshifts enhanced biofilm formation in the majority of Pseudomonas aeruginosa clinical and environmental isolates. A , biofilm biomass of 52 clinical P. aeruginosa isolates, P. aeruginosa PAO1, and four environmental isolates following culturing statically at RT and 37 °C. M1: ATCC 27853. M35: PA150663 . M54: PA152541 . B , dot plot summarizing the biofilm patterns of P. aeruginosa isolates at RT and 37 °C. Black : environmental isolates. The two dash lines represent the threshold of 2-fold. Blue : strains formed two-fold or more biofilm at RT relative to 37 °C. Red : strains formed two-fold or more biofilm at 37 °C relative to RT. See related for bar graph with statistical analysis. C , biofilm biomass of ATCC 27853, PAO1, PA150663 , and PA152541 at a temperature gradient from 15 °C to 42 °C at 48 h. D , biofilm formation of P. aeruginosa ATCC 27853 and PAO1 at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).
    Figure Legend Snippet: Temperature downshifts enhanced biofilm formation in the majority of Pseudomonas aeruginosa clinical and environmental isolates. A , biofilm biomass of 52 clinical P. aeruginosa isolates, P. aeruginosa PAO1, and four environmental isolates following culturing statically at RT and 37 °C. M1: ATCC 27853. M35: PA150663 . M54: PA152541 . B , dot plot summarizing the biofilm patterns of P. aeruginosa isolates at RT and 37 °C. Black : environmental isolates. The two dash lines represent the threshold of 2-fold. Blue : strains formed two-fold or more biofilm at RT relative to 37 °C. Red : strains formed two-fold or more biofilm at 37 °C relative to RT. See related for bar graph with statistical analysis. C , biofilm biomass of ATCC 27853, PAO1, PA150663 , and PA152541 at a temperature gradient from 15 °C to 42 °C at 48 h. D , biofilm formation of P. aeruginosa ATCC 27853 and PAO1 at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Techniques Used:

    Psl is primarily responsible for the enhanced biofilm formation at RT. A , SEM images of ATCC 27853 biofilm formed at RT ( upper ) and 37 °C ( lower ) on LB agar. Scale bar represents 1 μm. B , maximal biomass of ATCC 27853 and its isogenic Δ pslD, Δ pslF , and Δ pslFpslD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , relative activity of the chromosomal P psl - lux transcriptional reporter in ATCC 27853 collected at half maximal biofilm at RT and 37 °C, respectively. Data are represented as mean ± SD, n = 3 independent experiments. D , Western blot of PslD-3FLAG in ATCC 27853 cultured at RT and 37 °C. Mid-log phase cells were collected for the analysis ( C–D ), that is, static cultures harvested at 24 h at RT and 8 h at 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative Psl production of ATCC 27853 at RT and 37 °C as detected by immunoblot with Psl-specific antibody. Left panel, half peak biofilm time points (24h at RT and 8 h at 37 °C). Right panel, peak time points (48h at RT and 24 h at 37 °C). Data are represented as mean ± SD, n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). SEM, scanning electron microscopy.
    Figure Legend Snippet: Psl is primarily responsible for the enhanced biofilm formation at RT. A , SEM images of ATCC 27853 biofilm formed at RT ( upper ) and 37 °C ( lower ) on LB agar. Scale bar represents 1 μm. B , maximal biomass of ATCC 27853 and its isogenic Δ pslD, Δ pslF , and Δ pslFpslD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , relative activity of the chromosomal P psl - lux transcriptional reporter in ATCC 27853 collected at half maximal biofilm at RT and 37 °C, respectively. Data are represented as mean ± SD, n = 3 independent experiments. D , Western blot of PslD-3FLAG in ATCC 27853 cultured at RT and 37 °C. Mid-log phase cells were collected for the analysis ( C–D ), that is, static cultures harvested at 24 h at RT and 8 h at 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative Psl production of ATCC 27853 at RT and 37 °C as detected by immunoblot with Psl-specific antibody. Left panel, half peak biofilm time points (24h at RT and 8 h at 37 °C). Right panel, peak time points (48h at RT and 24 h at 37 °C). Data are represented as mean ± SD, n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). SEM, scanning electron microscopy.

    Techniques Used: Activity Assay, Western Blot, Cell Culture, Electron Microscopy

    Enhanced Psl production at RT was mediated by the c-di-GMP–FleQ pathway. A , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ fleQ , Δ rpoS , and Δ amrZ mutants at RT. Data are represented as mean ± SD, n = 3 independent experiments. B , relative activity of the chromosomal P psl - lux transcriptional fusion reporter collected at half maximal biofilm in ATCC 27853 and its isogenic Δ fleQ mutant at RT. Data are represented as mean ± SD, n = 3 independent experiments. C , intracellular c-di-GMP level at half maximal biofilm culture of ATCC 27853 at RT and 37 °C. Data are represented as mean ± SD, n = 2 independent experiments. D , dynamic biofilm formation of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate.
    Figure Legend Snippet: Enhanced Psl production at RT was mediated by the c-di-GMP–FleQ pathway. A , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ fleQ , Δ rpoS , and Δ amrZ mutants at RT. Data are represented as mean ± SD, n = 3 independent experiments. B , relative activity of the chromosomal P psl - lux transcriptional fusion reporter collected at half maximal biofilm in ATCC 27853 and its isogenic Δ fleQ mutant at RT. Data are represented as mean ± SD, n = 3 independent experiments. C , intracellular c-di-GMP level at half maximal biofilm culture of ATCC 27853 at RT and 37 °C. Data are represented as mean ± SD, n = 2 independent experiments. D , dynamic biofilm formation of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate.

    Techniques Used: Activity Assay, Mutagenesis

    The DGC SiaD is primarily responsible for the elevated c-di-GMP content and enhanced Psl production at RT in ATCC 27853. A , relative maximal biofilm biomass of 28 isogenic mutants relative to the ATCC 27853 WT at RT. Blue , genes identified in RNA-seq analysis. B , relative maximal biofilm formation of ATCC 27853 and its isogenic Δ siaD strain at RT and 37 °C. C , relative maximal biofilm formation of ATCC 27853, isogenic Δ siaD mutant, and Δ siaD mutant complemented with rhamnose-inducible expression of siaD at 48 h at RT. VC, the pJM253 vector control. D – F , relative activity of the chromosomal P psl - lux transcriptional fusion reporter ( D ), Psl level in biofilm matrix ( E ), and intracellular c-di-GMP level ( F ) in ATCC 27853 and its isogenic Δ siaD mutant at RT. Midlog phase static cells harvested at 24 h at RT were collected for the analysis in ( C–F ). Data are represented as mean ± SD. (A–E), n = 3 independent experiments. ( F ), n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.
    Figure Legend Snippet: The DGC SiaD is primarily responsible for the elevated c-di-GMP content and enhanced Psl production at RT in ATCC 27853. A , relative maximal biofilm biomass of 28 isogenic mutants relative to the ATCC 27853 WT at RT. Blue , genes identified in RNA-seq analysis. B , relative maximal biofilm formation of ATCC 27853 and its isogenic Δ siaD strain at RT and 37 °C. C , relative maximal biofilm formation of ATCC 27853, isogenic Δ siaD mutant, and Δ siaD mutant complemented with rhamnose-inducible expression of siaD at 48 h at RT. VC, the pJM253 vector control. D – F , relative activity of the chromosomal P psl - lux transcriptional fusion reporter ( D ), Psl level in biofilm matrix ( E ), and intracellular c-di-GMP level ( F ) in ATCC 27853 and its isogenic Δ siaD mutant at RT. Midlog phase static cells harvested at 24 h at RT were collected for the analysis in ( C–F ). Data are represented as mean ± SD. (A–E), n = 3 independent experiments. ( F ), n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Techniques Used: RNA Sequencing, Mutagenesis, Expressing, Plasmid Preparation, Control, Activity Assay

    Temperature downshift from 37 °C to RT induces biofilm formation in ATCC 27853 through the SiaABCD signaling module. A , schematic diagram of the SiaABCD signaling and functional pathway. B , relative maximal biomass of ATCC 27853 and its isogenic Δ siaA , Δ siaB, Δ siaC, and Δ siaD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , maximal biomass of ATCC 27853 supplemented with a vector control (VC) or rhamnose-inducible siaB at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. D , maximal biomass of ATCC 27853 and its isogenic mutants carrying siaC alleles encoding a phosphorylation-deficient (SiaC T68A ) or phosphomimetic (SiaC T68D ) mutant at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , phosphorylation status of SiaC-3FLAG at RT and 37 °C detected by Phos-tag SDS-PAGE. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test).
    Figure Legend Snippet: Temperature downshift from 37 °C to RT induces biofilm formation in ATCC 27853 through the SiaABCD signaling module. A , schematic diagram of the SiaABCD signaling and functional pathway. B , relative maximal biomass of ATCC 27853 and its isogenic Δ siaA , Δ siaB, Δ siaC, and Δ siaD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , maximal biomass of ATCC 27853 supplemented with a vector control (VC) or rhamnose-inducible siaB at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. D , maximal biomass of ATCC 27853 and its isogenic mutants carrying siaC alleles encoding a phosphorylation-deficient (SiaC T68A ) or phosphomimetic (SiaC T68D ) mutant at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , phosphorylation status of SiaC-3FLAG at RT and 37 °C detected by Phos-tag SDS-PAGE. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test).

    Techniques Used: Functional Assay, Plasmid Preparation, Control, Phospho-proteomics, Mutagenesis, SDS Page

    Temperature downshifts induce distinct membrane perturbations to activate the SiaABCD signaling module. A , 2D graph plotted from output signals of two channels of the PI-BactD probe, fluorescence increase (ΔS/S 0 ), and fluorescence radiometric changes (I 483 /I 375 ), in ATCC 27853 and PAO1 at RT and 37 °C. Each assay was conducted in six replicates. B , schematic diagram showing the domain structures of WT SiaA, WT CusS, and CusS-SiaA chimeric variant. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. Data are represented as mean ± SD, n = 3 independent experiments. D , upper panel, Log2 relative area of membrane lipid groups of ATCC 27853 and PAO1 as detected by lipidomics analysis (RT/37 °C). Orange and green : differentially changed in ATCC 27853 relative to PAO1 in response to temperature shift. Black and blue : synchronously changed in ATCC 27853 and PAO1 in response to temperature shift. Gray : similar levels at RT and 37 °C. Lower panel, a table summarizing the membrane perturbations upon temperature downshift and SDS treatment. E , dynamic biofilm formation of ATCC 27853 harboring either a vector control (VC) or rhamnose-inducible fabAB construct at RT and 37 °C. VC, the pJM253 vector control. Data are represented as mean ± SD, n = 3 independent experiments. F , pellicle formation of ATCC 27853 at RT and 37 °C collected at half-maximal biofilm stage, that is, 24h at RT and 8h at 37 °C, OD 600 ∼0.4. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).
    Figure Legend Snippet: Temperature downshifts induce distinct membrane perturbations to activate the SiaABCD signaling module. A , 2D graph plotted from output signals of two channels of the PI-BactD probe, fluorescence increase (ΔS/S 0 ), and fluorescence radiometric changes (I 483 /I 375 ), in ATCC 27853 and PAO1 at RT and 37 °C. Each assay was conducted in six replicates. B , schematic diagram showing the domain structures of WT SiaA, WT CusS, and CusS-SiaA chimeric variant. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. Data are represented as mean ± SD, n = 3 independent experiments. D , upper panel, Log2 relative area of membrane lipid groups of ATCC 27853 and PAO1 as detected by lipidomics analysis (RT/37 °C). Orange and green : differentially changed in ATCC 27853 relative to PAO1 in response to temperature shift. Black and blue : synchronously changed in ATCC 27853 and PAO1 in response to temperature shift. Gray : similar levels at RT and 37 °C. Lower panel, a table summarizing the membrane perturbations upon temperature downshift and SDS treatment. E , dynamic biofilm formation of ATCC 27853 harboring either a vector control (VC) or rhamnose-inducible fabAB construct at RT and 37 °C. VC, the pJM253 vector control. Data are represented as mean ± SD, n = 3 independent experiments. F , pellicle formation of ATCC 27853 at RT and 37 °C collected at half-maximal biofilm stage, that is, 24h at RT and 8h at 37 °C, OD 600 ∼0.4. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Techniques Used: Membrane, Fluorescence, Variant Assay, Mutagenesis, Plasmid Preparation, Control, Construct

    A model to illustrate the activation of SiaABCD and its mediated biofilm enhancement in ATCC 27853 in response to temperature downshifts. Temperature downshift from 37 °C to RT causes membrane perturbations in ATCC 27853, which induce SiaA phosphatase activity to promote the formation of SiaC–SiaD complex and DGC activity of SiaD. The activated Sia system regulates psl production via c-di-GMP–FleQ–Psl pathway. Increased Psl production enhanced biofilm formation of ATCC 27853 at RT through enhancing both surface attachment and biofilm matrix development. c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.
    Figure Legend Snippet: A model to illustrate the activation of SiaABCD and its mediated biofilm enhancement in ATCC 27853 in response to temperature downshifts. Temperature downshift from 37 °C to RT causes membrane perturbations in ATCC 27853, which induce SiaA phosphatase activity to promote the formation of SiaC–SiaD complex and DGC activity of SiaD. The activated Sia system regulates psl production via c-di-GMP–FleQ–Psl pathway. Increased Psl production enhanced biofilm formation of ATCC 27853 at RT through enhancing both surface attachment and biofilm matrix development. c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Techniques Used: Activation Assay, Membrane, Activity Assay

    Related Articles

    Mutagenesis:

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module
    Article Snippet: .. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. ..

    Variant Assay:

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module
    Article Snippet: .. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. ..



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    ATCC psa atcc 27853 wt
    Figure 5. Acb2 antagonizes trinucleotide- and dinucleotide-based CBASS immunity (A) Pseudomonas aeruginosa <t>BWHPSA011</t> <t>(Pa011)</t> type II-A CBASS and ATCC 27853 <t>(Pa278)</t> type III-C CBASS operons. (B) Pseudomonas aeruginosa PaMx33 and JBD67 phages acb2 gene annotated with residues essential for CDN (30,30-cGAMP) binding and CTN (cA3) binding. (C) Effect of PaMx33 Acb2 or its mutants on cA3-activated NucC effector protein function. After treatment with proteinase K, the released cA3 also showed the ability to activate the nuclease activity of NucC. The concentration of NucC, cA3, Acb2, and proteinase K is 10 nM, 5 nM, 50 nM, and 1 mM, respectively. N denotes nicked plasmid, SC denotes closed-circular supercoiled plasmid, and cut denotes fully digested DNA. (D) Plaque assays with JBD67Dacb2 phage spotted in 10-fold serial dilutions on PAO1 strains harboring an empty vector (E.V.) plasmid or JBD67 Acb2 variants. The PAO1 strains either contain no CBASS operon (CBASS), a chromosomally integrated Pa011 CBASS operon (PAO1Pa011), or a chromosomally integrated Pa278 CBASS operon (PAO1Pa278). These plaque assays were used to quantify the order of magnitude change in phage titer by comparing the number of spots (with plaques, or clearings if plaques were not visible) on the PAO1Pa011 or PAO1Pa278 CBASS-expressing strains divided by the PAO1 (CBASS) strain (n = 3). Basal expression of the Pa011 CBASS operon and 0.3 mM IPTG-inducible expression of the Pa278 CBASS operon is sufficient for phage targeting. Black ar- rowheads highlight significant CBASS-dependent reductions in phage titer. (E) Plaque assays with JBD67 phages spotted in 10-fold serial dilutions on PAO1 strains with and without CBASS. Pa278 CBASS was expressed from the pHERD30T (p30T) plasmid, while Pa011 CBASS was expressed from the chromosome. These plaque assays were used to quantify the order of magnitude change in phage titer (n = 3). Basal expression (i.e., no arabinose added) of the Pa278 CBASS operon is sufficient for phage targeting. Black arrowheads highlight significant CBASS-dependent reductions in phage titer.
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    Temperature downshifts enhanced biofilm formation in the majority of Pseudomonas aeruginosa clinical and environmental isolates. A , biofilm biomass of 52 clinical P. aeruginosa isolates, P. aeruginosa PAO1, and four environmental isolates following culturing statically at RT and 37 °C. M1: ATCC 27853. M35: PA150663 . M54: PA152541 . B , dot plot summarizing the biofilm patterns of P. aeruginosa isolates at RT and 37 °C. Black : environmental isolates. The two dash lines represent the threshold of 2-fold. Blue : strains formed two-fold or more biofilm at RT relative to 37 °C. Red : strains formed two-fold or more biofilm at 37 °C relative to RT. See related for bar graph with statistical analysis. C , biofilm biomass of ATCC 27853, PAO1, PA150663 , and PA152541 at a temperature gradient from 15 °C to 42 °C at 48 h. D , biofilm formation of P. aeruginosa ATCC 27853 and PAO1 at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: Temperature downshifts enhanced biofilm formation in the majority of Pseudomonas aeruginosa clinical and environmental isolates. A , biofilm biomass of 52 clinical P. aeruginosa isolates, P. aeruginosa PAO1, and four environmental isolates following culturing statically at RT and 37 °C. M1: ATCC 27853. M35: PA150663 . M54: PA152541 . B , dot plot summarizing the biofilm patterns of P. aeruginosa isolates at RT and 37 °C. Black : environmental isolates. The two dash lines represent the threshold of 2-fold. Blue : strains formed two-fold or more biofilm at RT relative to 37 °C. Red : strains formed two-fold or more biofilm at 37 °C relative to RT. See related for bar graph with statistical analysis. C , biofilm biomass of ATCC 27853, PAO1, PA150663 , and PA152541 at a temperature gradient from 15 °C to 42 °C at 48 h. D , biofilm formation of P. aeruginosa ATCC 27853 and PAO1 at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques:

    Psl is primarily responsible for the enhanced biofilm formation at RT. A , SEM images of ATCC 27853 biofilm formed at RT ( upper ) and 37 °C ( lower ) on LB agar. Scale bar represents 1 μm. B , maximal biomass of ATCC 27853 and its isogenic Δ pslD, Δ pslF , and Δ pslFpslD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , relative activity of the chromosomal P psl - lux transcriptional reporter in ATCC 27853 collected at half maximal biofilm at RT and 37 °C, respectively. Data are represented as mean ± SD, n = 3 independent experiments. D , Western blot of PslD-3FLAG in ATCC 27853 cultured at RT and 37 °C. Mid-log phase cells were collected for the analysis ( C–D ), that is, static cultures harvested at 24 h at RT and 8 h at 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative Psl production of ATCC 27853 at RT and 37 °C as detected by immunoblot with Psl-specific antibody. Left panel, half peak biofilm time points (24h at RT and 8 h at 37 °C). Right panel, peak time points (48h at RT and 24 h at 37 °C). Data are represented as mean ± SD, n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). SEM, scanning electron microscopy.

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: Psl is primarily responsible for the enhanced biofilm formation at RT. A , SEM images of ATCC 27853 biofilm formed at RT ( upper ) and 37 °C ( lower ) on LB agar. Scale bar represents 1 μm. B , maximal biomass of ATCC 27853 and its isogenic Δ pslD, Δ pslF , and Δ pslFpslD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , relative activity of the chromosomal P psl - lux transcriptional reporter in ATCC 27853 collected at half maximal biofilm at RT and 37 °C, respectively. Data are represented as mean ± SD, n = 3 independent experiments. D , Western blot of PslD-3FLAG in ATCC 27853 cultured at RT and 37 °C. Mid-log phase cells were collected for the analysis ( C–D ), that is, static cultures harvested at 24 h at RT and 8 h at 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative Psl production of ATCC 27853 at RT and 37 °C as detected by immunoblot with Psl-specific antibody. Left panel, half peak biofilm time points (24h at RT and 8 h at 37 °C). Right panel, peak time points (48h at RT and 24 h at 37 °C). Data are represented as mean ± SD, n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). SEM, scanning electron microscopy.

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: Activity Assay, Western Blot, Cell Culture, Electron Microscopy

    Enhanced Psl production at RT was mediated by the c-di-GMP–FleQ pathway. A , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ fleQ , Δ rpoS , and Δ amrZ mutants at RT. Data are represented as mean ± SD, n = 3 independent experiments. B , relative activity of the chromosomal P psl - lux transcriptional fusion reporter collected at half maximal biofilm in ATCC 27853 and its isogenic Δ fleQ mutant at RT. Data are represented as mean ± SD, n = 3 independent experiments. C , intracellular c-di-GMP level at half maximal biofilm culture of ATCC 27853 at RT and 37 °C. Data are represented as mean ± SD, n = 2 independent experiments. D , dynamic biofilm formation of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate.

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: Enhanced Psl production at RT was mediated by the c-di-GMP–FleQ pathway. A , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ fleQ , Δ rpoS , and Δ amrZ mutants at RT. Data are represented as mean ± SD, n = 3 independent experiments. B , relative activity of the chromosomal P psl - lux transcriptional fusion reporter collected at half maximal biofilm in ATCC 27853 and its isogenic Δ fleQ mutant at RT. Data are represented as mean ± SD, n = 3 independent experiments. C , intracellular c-di-GMP level at half maximal biofilm culture of ATCC 27853 at RT and 37 °C. Data are represented as mean ± SD, n = 2 independent experiments. D , dynamic biofilm formation of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , relative maximal biofilm biomass of ATCC 27853 and its isogenic Δ wspF strain at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate.

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: Activity Assay, Mutagenesis

    The DGC SiaD is primarily responsible for the elevated c-di-GMP content and enhanced Psl production at RT in ATCC 27853. A , relative maximal biofilm biomass of 28 isogenic mutants relative to the ATCC 27853 WT at RT. Blue , genes identified in RNA-seq analysis. B , relative maximal biofilm formation of ATCC 27853 and its isogenic Δ siaD strain at RT and 37 °C. C , relative maximal biofilm formation of ATCC 27853, isogenic Δ siaD mutant, and Δ siaD mutant complemented with rhamnose-inducible expression of siaD at 48 h at RT. VC, the pJM253 vector control. D – F , relative activity of the chromosomal P psl - lux transcriptional fusion reporter ( D ), Psl level in biofilm matrix ( E ), and intracellular c-di-GMP level ( F ) in ATCC 27853 and its isogenic Δ siaD mutant at RT. Midlog phase static cells harvested at 24 h at RT were collected for the analysis in ( C–F ). Data are represented as mean ± SD. (A–E), n = 3 independent experiments. ( F ), n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: The DGC SiaD is primarily responsible for the elevated c-di-GMP content and enhanced Psl production at RT in ATCC 27853. A , relative maximal biofilm biomass of 28 isogenic mutants relative to the ATCC 27853 WT at RT. Blue , genes identified in RNA-seq analysis. B , relative maximal biofilm formation of ATCC 27853 and its isogenic Δ siaD strain at RT and 37 °C. C , relative maximal biofilm formation of ATCC 27853, isogenic Δ siaD mutant, and Δ siaD mutant complemented with rhamnose-inducible expression of siaD at 48 h at RT. VC, the pJM253 vector control. D – F , relative activity of the chromosomal P psl - lux transcriptional fusion reporter ( D ), Psl level in biofilm matrix ( E ), and intracellular c-di-GMP level ( F ) in ATCC 27853 and its isogenic Δ siaD mutant at RT. Midlog phase static cells harvested at 24 h at RT were collected for the analysis in ( C–F ). Data are represented as mean ± SD. (A–E), n = 3 independent experiments. ( F ), n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: RNA Sequencing, Mutagenesis, Expressing, Plasmid Preparation, Control, Activity Assay

    Temperature downshift from 37 °C to RT induces biofilm formation in ATCC 27853 through the SiaABCD signaling module. A , schematic diagram of the SiaABCD signaling and functional pathway. B , relative maximal biomass of ATCC 27853 and its isogenic Δ siaA , Δ siaB, Δ siaC, and Δ siaD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , maximal biomass of ATCC 27853 supplemented with a vector control (VC) or rhamnose-inducible siaB at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. D , maximal biomass of ATCC 27853 and its isogenic mutants carrying siaC alleles encoding a phosphorylation-deficient (SiaC T68A ) or phosphomimetic (SiaC T68D ) mutant at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , phosphorylation status of SiaC-3FLAG at RT and 37 °C detected by Phos-tag SDS-PAGE. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test).

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: Temperature downshift from 37 °C to RT induces biofilm formation in ATCC 27853 through the SiaABCD signaling module. A , schematic diagram of the SiaABCD signaling and functional pathway. B , relative maximal biomass of ATCC 27853 and its isogenic Δ siaA , Δ siaB, Δ siaC, and Δ siaD mutants at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. C , maximal biomass of ATCC 27853 supplemented with a vector control (VC) or rhamnose-inducible siaB at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. D , maximal biomass of ATCC 27853 and its isogenic mutants carrying siaC alleles encoding a phosphorylation-deficient (SiaC T68A ) or phosphomimetic (SiaC T68D ) mutant at RT and 37 °C. Data are represented as mean ± SD, n = 3 independent experiments. E , phosphorylation status of SiaC-3FLAG at RT and 37 °C detected by Phos-tag SDS-PAGE. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test).

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: Functional Assay, Plasmid Preparation, Control, Phospho-proteomics, Mutagenesis, SDS Page

    Temperature downshifts induce distinct membrane perturbations to activate the SiaABCD signaling module. A , 2D graph plotted from output signals of two channels of the PI-BactD probe, fluorescence increase (ΔS/S 0 ), and fluorescence radiometric changes (I 483 /I 375 ), in ATCC 27853 and PAO1 at RT and 37 °C. Each assay was conducted in six replicates. B , schematic diagram showing the domain structures of WT SiaA, WT CusS, and CusS-SiaA chimeric variant. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. Data are represented as mean ± SD, n = 3 independent experiments. D , upper panel, Log2 relative area of membrane lipid groups of ATCC 27853 and PAO1 as detected by lipidomics analysis (RT/37 °C). Orange and green : differentially changed in ATCC 27853 relative to PAO1 in response to temperature shift. Black and blue : synchronously changed in ATCC 27853 and PAO1 in response to temperature shift. Gray : similar levels at RT and 37 °C. Lower panel, a table summarizing the membrane perturbations upon temperature downshift and SDS treatment. E , dynamic biofilm formation of ATCC 27853 harboring either a vector control (VC) or rhamnose-inducible fabAB construct at RT and 37 °C. VC, the pJM253 vector control. Data are represented as mean ± SD, n = 3 independent experiments. F , pellicle formation of ATCC 27853 at RT and 37 °C collected at half-maximal biofilm stage, that is, 24h at RT and 8h at 37 °C, OD 600 ∼0.4. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: Temperature downshifts induce distinct membrane perturbations to activate the SiaABCD signaling module. A , 2D graph plotted from output signals of two channels of the PI-BactD probe, fluorescence increase (ΔS/S 0 ), and fluorescence radiometric changes (I 483 /I 375 ), in ATCC 27853 and PAO1 at RT and 37 °C. Each assay was conducted in six replicates. B , schematic diagram showing the domain structures of WT SiaA, WT CusS, and CusS-SiaA chimeric variant. C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT. Data are represented as mean ± SD, n = 3 independent experiments. D , upper panel, Log2 relative area of membrane lipid groups of ATCC 27853 and PAO1 as detected by lipidomics analysis (RT/37 °C). Orange and green : differentially changed in ATCC 27853 relative to PAO1 in response to temperature shift. Black and blue : synchronously changed in ATCC 27853 and PAO1 in response to temperature shift. Gray : similar levels at RT and 37 °C. Lower panel, a table summarizing the membrane perturbations upon temperature downshift and SDS treatment. E , dynamic biofilm formation of ATCC 27853 harboring either a vector control (VC) or rhamnose-inducible fabAB construct at RT and 37 °C. VC, the pJM253 vector control. Data are represented as mean ± SD, n = 3 independent experiments. F , pellicle formation of ATCC 27853 at RT and 37 °C collected at half-maximal biofilm stage, that is, 24h at RT and 8h at 37 °C, OD 600 ∼0.4. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01 (based on Student’s t test).

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: Membrane, Fluorescence, Variant Assay, Mutagenesis, Plasmid Preparation, Control, Construct

    A model to illustrate the activation of SiaABCD and its mediated biofilm enhancement in ATCC 27853 in response to temperature downshifts. Temperature downshift from 37 °C to RT causes membrane perturbations in ATCC 27853, which induce SiaA phosphatase activity to promote the formation of SiaC–SiaD complex and DGC activity of SiaD. The activated Sia system regulates psl production via c-di-GMP–FleQ–Psl pathway. Increased Psl production enhanced biofilm formation of ATCC 27853 at RT through enhancing both surface attachment and biofilm matrix development. c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Journal: The Journal of Biological Chemistry

    Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module

    doi: 10.1016/j.jbc.2025.111086

    Figure Lengend Snippet: A model to illustrate the activation of SiaABCD and its mediated biofilm enhancement in ATCC 27853 in response to temperature downshifts. Temperature downshift from 37 °C to RT causes membrane perturbations in ATCC 27853, which induce SiaA phosphatase activity to promote the formation of SiaC–SiaD complex and DGC activity of SiaD. The activated Sia system regulates psl production via c-di-GMP–FleQ–Psl pathway. Increased Psl production enhanced biofilm formation of ATCC 27853 at RT through enhancing both surface attachment and biofilm matrix development. c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.

    Article Snippet: C , maximal biomass of ATCC 27853 WT strain, Δ siaA mutant, and Δ siaA mutant carrying chromosomally integrated CusS-SiaA chimeric variant at RT.

    Techniques: Activation Assay, Membrane, Activity Assay

    Figure 5. Acb2 antagonizes trinucleotide- and dinucleotide-based CBASS immunity (A) Pseudomonas aeruginosa BWHPSA011 (Pa011) type II-A CBASS and ATCC 27853 (Pa278) type III-C CBASS operons. (B) Pseudomonas aeruginosa PaMx33 and JBD67 phages acb2 gene annotated with residues essential for CDN (30,30-cGAMP) binding and CTN (cA3) binding. (C) Effect of PaMx33 Acb2 or its mutants on cA3-activated NucC effector protein function. After treatment with proteinase K, the released cA3 also showed the ability to activate the nuclease activity of NucC. The concentration of NucC, cA3, Acb2, and proteinase K is 10 nM, 5 nM, 50 nM, and 1 mM, respectively. N denotes nicked plasmid, SC denotes closed-circular supercoiled plasmid, and cut denotes fully digested DNA. (D) Plaque assays with JBD67Dacb2 phage spotted in 10-fold serial dilutions on PAO1 strains harboring an empty vector (E.V.) plasmid or JBD67 Acb2 variants. The PAO1 strains either contain no CBASS operon (CBASS), a chromosomally integrated Pa011 CBASS operon (PAO1Pa011), or a chromosomally integrated Pa278 CBASS operon (PAO1Pa278). These plaque assays were used to quantify the order of magnitude change in phage titer by comparing the number of spots (with plaques, or clearings if plaques were not visible) on the PAO1Pa011 or PAO1Pa278 CBASS-expressing strains divided by the PAO1 (CBASS) strain (n = 3). Basal expression of the Pa011 CBASS operon and 0.3 mM IPTG-inducible expression of the Pa278 CBASS operon is sufficient for phage targeting. Black ar- rowheads highlight significant CBASS-dependent reductions in phage titer. (E) Plaque assays with JBD67 phages spotted in 10-fold serial dilutions on PAO1 strains with and without CBASS. Pa278 CBASS was expressed from the pHERD30T (p30T) plasmid, while Pa011 CBASS was expressed from the chromosome. These plaque assays were used to quantify the order of magnitude change in phage titer (n = 3). Basal expression (i.e., no arabinose added) of the Pa278 CBASS operon is sufficient for phage targeting. Black arrowheads highlight significant CBASS-dependent reductions in phage titer.

    Journal: Molecular cell

    Article Title: Phage anti-CBASS protein simultaneously sequesters cyclic trinucleotides and dinucleotides.

    doi: 10.1016/j.molcel.2023.11.026

    Figure Lengend Snippet: Figure 5. Acb2 antagonizes trinucleotide- and dinucleotide-based CBASS immunity (A) Pseudomonas aeruginosa BWHPSA011 (Pa011) type II-A CBASS and ATCC 27853 (Pa278) type III-C CBASS operons. (B) Pseudomonas aeruginosa PaMx33 and JBD67 phages acb2 gene annotated with residues essential for CDN (30,30-cGAMP) binding and CTN (cA3) binding. (C) Effect of PaMx33 Acb2 or its mutants on cA3-activated NucC effector protein function. After treatment with proteinase K, the released cA3 also showed the ability to activate the nuclease activity of NucC. The concentration of NucC, cA3, Acb2, and proteinase K is 10 nM, 5 nM, 50 nM, and 1 mM, respectively. N denotes nicked plasmid, SC denotes closed-circular supercoiled plasmid, and cut denotes fully digested DNA. (D) Plaque assays with JBD67Dacb2 phage spotted in 10-fold serial dilutions on PAO1 strains harboring an empty vector (E.V.) plasmid or JBD67 Acb2 variants. The PAO1 strains either contain no CBASS operon (CBASS), a chromosomally integrated Pa011 CBASS operon (PAO1Pa011), or a chromosomally integrated Pa278 CBASS operon (PAO1Pa278). These plaque assays were used to quantify the order of magnitude change in phage titer by comparing the number of spots (with plaques, or clearings if plaques were not visible) on the PAO1Pa011 or PAO1Pa278 CBASS-expressing strains divided by the PAO1 (CBASS) strain (n = 3). Basal expression of the Pa011 CBASS operon and 0.3 mM IPTG-inducible expression of the Pa278 CBASS operon is sufficient for phage targeting. Black ar- rowheads highlight significant CBASS-dependent reductions in phage titer. (E) Plaque assays with JBD67 phages spotted in 10-fold serial dilutions on PAO1 strains with and without CBASS. Pa278 CBASS was expressed from the pHERD30T (p30T) plasmid, while Pa011 CBASS was expressed from the chromosome. These plaque assays were used to quantify the order of magnitude change in phage titer (n = 3). Basal expression (i.e., no arabinose added) of the Pa278 CBASS operon is sufficient for phage targeting. Black arrowheads highlight significant CBASS-dependent reductions in phage titer.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains P. aeruginosa BWHPSA011 (Pa011) WT Deborah Hung Lab NCBI: NZ_AXQR01000000.1 P. aeruginosa ATCC 27853 (Pa278) WT ATCC NCBI: CP015177.1 P. aeruginosa PAO1 WT Joe Bondy-Denomy Lab NCBI: NC_002516.2 PAO1Pa011 (PAO1 attTn7::Pa011 Type II-A CBASS) Joe Bondy-Denomy Lab16 N/A PAO1Pa278 (PAO1 attTn7::Pa278 Type III-C CBASS) This study N/A JBD67 Alan Davidson Lab30 NCBI: NC_042135.1 JBD67Dacb2 (Removal of 23005-23232 bp orf24 (acb2)) Joe Bondy-Denomy Lab16 N/A JBD67 acb2K26A This study N/A JBD67 acb2R82A This study N/A JBD67 acb2R82A/T89A This study N/A JBD67 acb2K26A This study N/A Experimental models: Cell lines HEK293T Dual-Null Invivogen Cat#: 293d-null Recombinant DNA pET28a-His6-SUMO- PaMx33-Acb2 This study N/A pET28a-His6-SUMO- PaMx33-Acb2 Y11A This study N/A pET28a-His6-SUMO- PaMx33-Acb2 K26A This study N/A pET28a-His6-SUMO- PaMx33-Acb2 T74A This study N/A pET28a-His6-SUMO- PaMx33-Acb2 R67A This study N/A pET28a-His6-SUMO- JBD67-Acb2 This study N/A pET28a-His6-CHI14-Acb2 This study N/A pET28a-His6-CHI14-Acb2 E76T This study N/A pET28a-His6-CHI14- Acb2 E76T/R67P This study N/A pET28a-His6-SUMO-T4-Acb2 This study N/A pET28a-His6-SUMO-T4-Acb2 D61R This study N/A pUC18-mini-Tn7-LAC (pTn7) Choi and Schweizer31 N/A pTn7-Pa278 Type III-C CBASS (6283357-6286926) This study N/A pNTS3 Choi et al.32 N/A pMQ30 Shanks et al.33 N/A pMQ30-HDR-JBD67 Acb2 WT This study N/A pMQ30-HDR-JBD67 Acb2 K26A This study N/A pMQ30-HDR-JBD67 Acb2 R82A This study N/A pMQ30-HDR-JBD67 Acb2 T89A This study N/A pHERD30T (p30T) Qiu et al.34 N/A p30T-JBD67 WT Acb2 This study N/A p30T-JBD67 Y11A Acb2 This study N/A p30T-JBD67 K26A Acb2 This study N/A p30T-JBD67 R82A Acb2 This study N/A p30T-JBD67 T89A Acb2 This study N/A p30T-Pa27853 Type III-C CBASS (6283357-6286926) This study N/A pcDNA3 Lingyin Li Lab N/A (Continued on next page) e1 Molecular Cell 84, 375–385.e1–e7, January 18, 2024

    Techniques: Binding Assay, Activity Assay, Concentration Assay, Plasmid Preparation, Expressing

    MIC of Ng strains (A) and Gram-negative pathogens and gut microorganisms (B). (A) WHO F does not carry any antimicrobial resistance elements and is considered an antibiotic susceptible strain. WHO G-M strains are resistant to quinolones. WHO K-O strains carry penA , ponA , porB1 , and mtrR mutations associated with decreased susceptibility to cephalosporins. ( <xref ref-type= Unemo et al., 2016 ). N. meningitidis (NMSB) ATCC 13090 is the suggested reference strain for serogroup B ( Clinical and Laboratory Standards Institute, 2004 ). (B) This panel was constructed to test for inhibition of Gram-negative pathogens and normally occurring intestinal organisms, where WT = wild type, MDR = multiple drug resistant, and QuinR = quinolone-resistant strains. The Nm , Gram-negative, and normal gut organism strains were obtained from the American Type Culture Collection (ATCC), or were kindly provided by MicroMyx, LLC, (MMX), Kalamazoo, MI, or the laboratory of Lynn Zechiedrich (LZ), Baylor College of Medicine, Houston, Texas. PTC-compound susceptibility testing was performed in accordance with the Clinical and Laboratory Standards Institute (CLSI) M07-A9 guideline ( Clinical and Laboratory Standards Institute, 2012 ). Genus names: Acinetobacter (A), Klebsiella (K), Pseudomonas (P), Staphylococcus (S), Yersinia (Y)." width="100%" height="100%">

    Journal: eLife

    Article Title: Ribonucleotide reductase, a novel drug target for gonorrhea

    doi: 10.7554/eLife.67447

    Figure Lengend Snippet: MIC of Ng strains (A) and Gram-negative pathogens and gut microorganisms (B). (A) WHO F does not carry any antimicrobial resistance elements and is considered an antibiotic susceptible strain. WHO G-M strains are resistant to quinolones. WHO K-O strains carry penA , ponA , porB1 , and mtrR mutations associated with decreased susceptibility to cephalosporins. ( Unemo et al., 2016 ). N. meningitidis (NMSB) ATCC 13090 is the suggested reference strain for serogroup B ( Clinical and Laboratory Standards Institute, 2004 ). (B) This panel was constructed to test for inhibition of Gram-negative pathogens and normally occurring intestinal organisms, where WT = wild type, MDR = multiple drug resistant, and QuinR = quinolone-resistant strains. The Nm , Gram-negative, and normal gut organism strains were obtained from the American Type Culture Collection (ATCC), or were kindly provided by MicroMyx, LLC, (MMX), Kalamazoo, MI, or the laboratory of Lynn Zechiedrich (LZ), Baylor College of Medicine, Houston, Texas. PTC-compound susceptibility testing was performed in accordance with the Clinical and Laboratory Standards Institute (CLSI) M07-A9 guideline ( Clinical and Laboratory Standards Institute, 2012 ). Genus names: Acinetobacter (A), Klebsiella (K), Pseudomonas (P), Staphylococcus (S), Yersinia (Y).

    Article Snippet: P. aeruginosa WT , ATCC 27853 , 12.5 , > 62.5 , > 62.5.

    Techniques: Construct, Inhibition