A conserved mycobacterial nucleomodulin hijacks the host COMPASS complex to reprogram pro-inflammatory transcription and promote intracellular survival
- National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, China
- Hubei Hongshan Laboratory, Huazhong Agricultural University, China
- Hubei Jiangxia Laboratory, China
- National Professional Laboratory for Animal Tuberculosis, Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, China
Figures
Figure 1 with 1 supplement
Identification of conserved nucleomodulins in mycobacteria through functional screening.
(A) Schematic representation of the bioinformatic pipeline for identifying conserved nucleomodulins in Mycobacterium species. Genomic sequences from M. tuberculosis H37Rv, M. tuberculosis H37Ra, M. avium, M. marinum, and M. bovis BCG were analyzed. Signal peptides were predicted using SignalP 5.0 (D-score ≥0.5), non-classical secretion signals were identified using SecretomeP 2.0 (NN score ≥0.9), and nuclear localization signals (NLS) were predicted using cNLS Mapper (score ≥2.0). (B) Classification of conserved cellular nucleomodulins. Secreted proteins were categorized based on the presence or absence of predicted NLS motifs. (C) Subcellular localization of enhanced green fluorescent protein (EGFP)-tagged candidate nucleomodulins. (left) Confocal microscopy images of seven Tat/SPI proteins fused to EGFP (green), nuclei were stained with DAPI (blue). (right) Quantification of nuclear EGFP fluorescence intensity. Scale bar, 10 µm. Data are presented as mean ± SD (n=25 cells/group).
Figure 1—figure supplement 1
Comparative analysis of classical and non-classical secreted proteins in mycobacterial species.
(A) Venn diagram showing the distribution of predicted classical secreted proteins in four Mycobacterium species: M. tuberculosis H37Rv (Mtu) and M. tuberculosis H37Ra (Mra), M. bovis BCG (Mbb), M. marinum (Mmar), and M. avium (Mav). Proteins were predicted using SignalP 5.0, with a signal peptide score (D-score) ≥0.5. A total of 125 proteins were conserved across all species. (B) Venn diagram showing the distribution of predicted non-classical secreted proteins across the same Mycobacterium species, predicted using SecretomeP 2.0 with a neural network (NN) score ≥0.9. Ten proteins were conserved across all species. Bar graphs (left: classical, right: non-classical) summarize the total number of predicted secreted proteins per species.
Figure 2 with 1 supplement
Hypothetical protein MgdE enters the host nucleus via dual nuclear localization signal (NLS).
(A) Domain architecture of MgdE. Schematic representation of MgdE with the annotated functional domains, including a Tat signal peptide (1–37 aa, twin-arginine translocation motif), a GDSL-like lipase/acylhydrolase catalytic domain (74–258 aa), and two nuclear localization signals (NLS1: 108–111 aa, NLS2: 300–305 aa). (B) Phylogenetic and structural conservation of MgdE. (left) Neighbor-joining phylogenetic tree of MgdE homologs across Mycobacterium species (1000 bootstrap replicates, values ≥50% shown). (right) Clustal Omega sequence alignment highlighting conserved residues (≥90% identity, red). (C) Subcellular localization of EGFP-tagged wild-type MgdE and its NLS-deletion mutants (MgdEΔNLS1, MgdEΔNLS2, and MgdEΔNLS1-2). (left) Schematic representation of EGFP-tagged constructs. (right) Representative confocal microscopy images of HEK293T cells transfected with the indicated constructs for 36 hr. Enhanced green fluorescent protein (EGFP) fluorescence (green) and nuclear staining with DAPI (blue) were visualized using FLUOVIEW software. Scale bar, 10 μm. Images were acquired using a 100x oil immersion objective (NA = 1.4). (D) Quantification of nuclear EGFP intensity in cells expressing wild-type or mutant MgdE constructs. Data are presented as mean ± SD (n=15 cells). Statistical significance was assessed by two-tailed unpaired Student’s t-tests (***p<0.001). (E) Western blot analysis of nuclear and cytoplasmic fractions from HEK293T cells transfected with wild-type MgdE and its NLS-deletion mutants. The empty EGFP vector was used as a negative control. EGFP and MgdE-EGFP fusion proteins were detected using an anti-GFP antibody. Histone H3 and β-actin served as nuclear and cytoplasmic markers, respectively. (F) Nuclear localization of MgdE during infection. THP-1 macrophages were infected with recombinant M. bovis BCG strains expressing Flag-tagged wild-type MgdE or NLS-deletion mutants (MgdEΔNLS1, MgdEΔNLS2, and MgdEΔNLS1-2) for 24 hr. Subcellular fractionation was performed, and cytoplasmic and nuclear fractions were analyzed by immunoblotting using anti-Flag antibodies. Histone H3 and β-actin were used as nuclear and cytoplasmic markers, respectively.
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Figure 2—source data 1
Original files for western blot analysis displayed in panel E.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-data1-v1.zip
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Figure 2—source data 2
Original western blots for panel E, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-data2-v1.zip
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Figure 2—source data 3
Original files for western blot analysis displayed in panel F.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-data3-v1.zip
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Figure 2—source data 4
Original western blots for panel F, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-data4-v1.zip
Figure 2—figure supplement 1
Subnuclear localization of MgdE-enhanced green fluorescent protein (EGFP).
(A) Immunoblot analysis of bacterial lysates and culture supernatants from M. bovis BCG strains expressing C-terminally Flag-tagged MgdE. Ag85B and GlpX were detected using anti-Ag85B and anti-GlpX antibodies, serving as positive and negative controls for protein secretion, respectively. MgdE-Flag was detected using an anti-Flag antibody. (B) Confocal microscopy was used to assess the nuclear distribution of MgdE-EGFP at various time points post-transfection. Nuclei were stained with DAPI (blue), MgdE-EGFP is shown in green. Scale bar: 10 μm. Images were acquired using a 100x oil immersion objective (NA = 1.4). (C) Quantification of nuclear EGFP intensity in cells expressing wild-type or mutant MgdE constructs. Data are presented as mean ± SD (n=12 cells). (D) Quantitative RT-PCR analysis of mgdE mRNA expression in HEK293T cells at different time points post-transfection (4-48 hr). Data represent the transcriptional level of mgdE relative to HPRT. (E) Western blot analysis of nuclear fractions at different time points post-transfection (4-48 hr), showing time-dependent nuclear accumulation of MgdE-EGFP. Histone H3 was used as a loading control for nuclear proteins. The lower panel shows quantification of nuclear MgdE-EGFP levels normalized to Histone H3. (F) Quantification of the nuclear and cytoplasmic distribution of EGFP, wild-type MgdE, and its NLS-deletion mutants based on the Western blot results shown in (Figure 2E). Data represent mean ± SD of three independent biological replicates. Statistical significance was determined using two-tailed unpaired Student’s t-tests, *p<0.05, **p<0.01, ***p<0.001.
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Figure 2—figure supplement 1—source data 1
Original files for western blot analysis displayed in panel A.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-figsupp1-data1-v1.zip
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Figure 2—figure supplement 1—source data 2
Original western blots for panel A, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-figsupp1-data2-v1.zip
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Figure 2—figure supplement 1—source data 3
Original files for western blot analysis displayed in panel E.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-figsupp1-data3-v1.zip
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Figure 2—figure supplement 1—source data 4
Original western blots for panel E, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig2-figsupp1-data4-v1.zip
Figure 3 with 1 supplement
Nuclear localization of MgdE facilitates mycobacterial intracellular survival in macrophages.
(A–B) Intracellular survival of M. bovis BCG strains. THP-1 human macrophages (A) and RAW264.7 murine macrophages (B) were infected (MOI = 10) with wild-type BCG (WT), MgdE deletion mutant (ΔMgdE), complemented strain (Comp-MgdE), or NLS-deficient complement (Comp-MgdEΔNLS1-2). Bacterial survival was assessed by colony-forming unit (CFU) enumeration at 2, 24, 48, and 72 hr post-infection. (C–H) Cytokine expression in the infected THP-1 cells. qRT-PCR analysis of cytokine mRNA levels in THP-1 cells infected with WT or ΔMgdE strains for 4-24 hr. Target genes include IL1B (C), IL6 (D), IL10 (E), CSF1 (F), CSF2 (G), and CSF3 (H). Data represent mean ± SD of three independent biological replicates. Statistical significance was determined using two-way ANOVA or two-tailed unpaired Student’s t-tests, *p<0.05, **p<0.01, and ***p<0.001.
Figure 3—figure supplement 1
Deletion of the nuclear localization signal of MgdE does not affect the growth of M.
bovis BCG strains. (A–B) Construction and validation of the MgdE-deleted strain of M. bovis BCG. (A) Schematic diagram of the homologous recombination strategy used to delete mgdE from the M. bovis BCG genome. (B) Wild-type and mutant strains were used as templates to amplify the mgdE gene (600 bp upstream-600 bp downstream) by PCR. Lanes 1 and 3: wild-type genomic DNA, lanes 2 and 4: ΔmgdE genomic DNA. (C) Growth curve analysis of M. bovis BCG strains. Growth of BCG strains, including wild-type BCG (WT), MgdE-deleted (ΔMgdE), MgdE-complemented (Comp-MgdE), and nuclear localization signal (NLS)-deleted complemented (Comp-MgdEΔNLS1-2) strains, was measured in 7H9 medium. Data represent mean ± SD of three independent biological replicates.
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Figure 3—figure supplement 1—source data 1
Original files for western blot analysis displayed in panel B.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig3-figsupp1-data1-v1.zip
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Figure 3—figure supplement 1—source data 2
Original western blots for panel B, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig3-figsupp1-data2-v1.zip
Figure 4 with 1 supplement
MgdE directly interacts with ASH2L and WDR5, core components of the host COMPASS complex.
(A) Yeast cells were co-transformed with bait (pGBKT7) and prey (pGADT7) plasmids expressing wild-type MgdE and human COMPASS components (ASH2L, WDR5, RbBP5, and DPY30). Growth was monitored on non-selective (-Leu/-Trp, left) and selective (-Leu/-Trp/-Ade/-His+200 ng/μL aureobasidin A, right) media. Controls: CK− (pGBKT7-lam+pGADT7 T, negative), CK+ (pGBKT7-p53+pGADT7 T, positive). (B–C) Cells were co-transfected with Flag-MgdE and HA-tagged ASH2L, WDR5, or RbBP5 (1:1 molar ratio). At 36 hr post-transfection, the lysates were immunoprecipitated using (B) anti-HA or (C) anti-Flag antibodies, followed by immunoblotting with anti-HA, anti-Flag, and anti-GAPDH (loading control). The input lanes represent 5% of the total lysate.
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Figure 4—source data 1
Original files for western blot analysis displayed in panel B.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-data1-v1.zip
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Figure 4—source data 2
Original western blots for panel B, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-data2-v1.zip
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Figure 4—source data 3
Original files for western blot analysis displayed in panel C.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-data3-v1.zip
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Figure 4—source data 4
Original western blots for panel C, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-data4-v1.zip
Figure 4—figure supplement 1
MgdE interacts with COMPASS complex subunits.
(A) Predicted binding affinities between MgdE and COMPASS core subunits. The predicted local distance difference test (pLDDT) scores calculated using AlphaFold 2.2.0 for the interactions between MgdE and the COMPASS subunits were as follows: ASH2L (pLDDT = 0.47), RbBP5 (pLDDT = 0.30), WDR5 (pLDDT = 0.77), and DPY30 (pLDDT = 0.62). Confidence levels are categorized as follows: High confidence: pLDDT ≥0.7 (strong predicted binding), Medium confidence: 0.5 ≤ pLDDT < 0.7 (moderate binding), Low confidence: pLDDT <0.5 (weak predicted binding). (B) Structural modeling of MgdE-COMPASS interactions. AlphaFold-generated models of the simulated binding interfaces are shown, with MgdE highlighted in red and COMPASS subunits (ASH2L, WDR5, RbBP5, and DPY30) shown in gray. (C) Co-IP analysis of MgdE mutants with ASH2L. HEK293T cells were co-transfected with Flag-tagged MgdE mutants and HA-tagged ASH2L (1:1 molar ratio). Co-IP was performed using anti-HA antibody-bound protein A/G beads, and immunoprecipitated complexes were analyzed by immunoblotting with anti-Flag and anti-HA antibodies.
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Figure 4—figure supplement 1—source data 1
Original files for western blot analysis displayed in panel C.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-figsupp1-data1-v1.zip
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Figure 4—figure supplement 1—source data 2
Original western blots for panel C, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig4-figsupp1-data2-v1.zip
Figure 5
The conserved residues D224 and H247 mediate the binding ability of MgdE to WDR5.
(A) Y2H assay identifying interactions between MgdE mutants and COMPASS complex subunits. Yeast cells were co-transformed with bait (pGBKT7) and prey (pGADT7) plasmids expressing wild-type or mutant MgdE and human COMPASS subunits (ASH2L, WDR5, RbBP5, and DPY30). Growth was assessed on non-selective (-Leu/-Trp, left) and selective (-Leu/-Trp/-Ade/-His+200 ng/μL aureobasidin A, right) media. Controls: CK− (pGBKT7-lam+pGADT7 T, negative) and CK+ (pGBKT7-p53+pGADT7 T, positive). (B) Co-IP analysis of MgdE mutants with WDR5. HEK293T cells were co-transfected with Flag-tagged MgdE mutants and HA-tagged WDR5 (1:1 molar ratio). Complexes were immunoprecipitated using anti-HA antibody and protein A/G beads, followed by immunoblotting with anti-Flag and anti-HA antibodies. (C) Nuclear distribution of wild-type and mutants MgdE. Confocal microscopy of HEK293T cells expressing wild-type or D224AH247A MgdE-EGFP at 12 and 24 hr post-transfection (hpt). Nuclear foci were visualized by enhanced green fluorescent protein (EGFP) (green) and DAPI (blue) staining. Scale bar, 10 µm. Images were acquired with a 100x oil immersion objective (NA = 1.4). (D) Immunoblot analysis of H3K4me3 levels. HEK293T cells expressing wild-type or D224AH247A mutant MgdE were analyzed for changes in H3K4me3 levels over 0-24 hr post-transfection. Histone H3 was used as a loading control. Data represent mean ± SD of three independent biological replicates. Statistical significance was determined using two-tailed unpaired Student’s t-tests, *p<0.05, **p<0.01, ***p<0.001.
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Figure 5—source data 1
Original files for western blot analysis displayed in panel B.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig5-data1-v1.zip
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Figure 5—source data 2
Original western blots for panel B, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig5-data2-v1.zip
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Figure 5—source data 3
Original files for western blot analysis displayed in panel D.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig5-data3-v1.zip
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Figure 5—source data 4
Original western blots for panel D, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/107677/elife-107677-fig5-data4-v1.zip
Figure 6 with 1 supplement
MgdE suppresses host inflammatory responses probably by inhibition of COMPASS complex-mediated H3K4 methylation.
(A) Volcano plot of DEGs. DEGs between MgdE-deleted strain (ΔMgdE) and wild-type BCG (WT) were visualized in a volcano plot. Genes with |log2-fold change|≥1 and p<0.05 were considered significant. The x-axis represents log2fold change, and the y-axis shows -log10(p-value). (B) Gene ontology (GO) enrichment analysis of DEGs. GO analysis revealed the significant enrichment of immune and inflammatory processes in ΔMgdE-infected macrophages compared to that in WT-infected cells, including positive regulation of response to external stimulus (GO:0032103), response to molecule of bacterial origin (GO:0002237), response to virus (GO:0009615), response to lipopolysaccharide (GO:0032496), leukocyte migration (GO:0050900), viral process (GO:0016032), regulation of response to biotic stimulus (GO:0002831), T cell activation (GO:0042110), and mononuclear cell differentiation (GO:1903131). (C) KEGG pathway enrichment analysis of DEGs. Chord diagram of KEGG pathway enrichment analysis showing signaling pathways that are significantly enriched in ΔMgdE. (D) Heatmap of the inflammatory gene expression. Heatmap depicting log2-fold change levels of inflammatory genes involved in the JAK-STAT and cytokine signaling pathways. Upregulated and downregulated genes in ΔMgdE are shown in green and red, respectively. The data were Z-score normalized. (E–G) qRT-PCR analysis of cytokine mRNA levels in THP-1 cells infected with WT, ΔMgdE, MgdE-complemented (Comp-MgdE), and nuclear localization signal (NLS)-deleted complement (Comp-MgdEΔNLS1-2) at 24 hr post-infection. Cytokines analyzed include IL1B (E), IL6 (F), and IL10 (G). Data represent mean ± SD of three independent biological replicates. Statistical significance was determined using two-tailed unpaired Student’s t-tests, **p<0.01, ***p<0.001.
Figure 6—figure supplement 1
MgdE suppresses cellular inflammatory responses during M. bovis BCG infection.
(A) Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. KEGG pathway analysis identified significantly enriched pathways in THP-1 cells infected with the ΔMgdE strain compared to those infected with wild-type BCG (WT). DEGs were predominantly associated with immune and signaling pathways, including thyroid hormone synthesis (hsa04918), AMPK signaling pathway (hsa04152), PPAR signaling pathway (hsa03320), apelin signaling pathway (hsa04371), Cytokine-cytokine receptor interaction (hsa04060), JAK-STAT signaling pathway (hsa04630), and hematopoietic cell lineage (hsa04640). (B) Elevated inflammatory gene expression in ΔMgdE-infected cells. Key upregulated inflammatory genes in ΔMgdE- vs. WT-infected cells are highlighted. (C) Functional enrichment analysis of upregulated genes. Analysis using STRING and Cytoscape revealed regulation of inflammatory responses as a top-enriched biological process (p<0.05).
Figure 7 with 1 supplement
Nuclear localization of MgdE is essential for mycobacterial survival in mice.
(A) Bacterial burden in the lungs of the infected mice. C57BL/6 mice (n=6/group) maintained under specific pathogen-free (SPF) conditions were intratracheally infected with 1.0×107 colony-forming units (CFU) of M. bovis BCG strains, including wild-type (WT), MgdE-deleted (ΔMgdE), MgdE-complemented (Comp-MgdE), and nuclear localization signals (NLS)-deleted complement (Comp-MgdEΔNLS1-2). Lung bacterial loads were quantified using CFU assays at 0, 14, 21, 28, and 56 days post-infection. The data were obtained from a single experiment. (B) Hematoxylin and eosin (H&E)-stained lung sections from infected mice (as in A) revealed granulomatous inflammation. Scale bars: 200 μm. (C-D) Pro-inflammatory cytokine expression in mice spleen. qRT-PCR analysis of cytokine mRNA levels of Il1b (C) and Il6 (D) in spleen tissues from infected mice (n=6/group) at 2 and 28 days post-infection. Data are presented as mean ± SD from six biologically independent experiments. Statistical significance was determined using two-way ANOVA, *p<0.05, **p<0.01, ***p<0.001. (E) Mechanistic model showing how mycobacterial nucleomodulin MgdE hijacks the COMPASS complex to suppress H3K4me3 and promote immune evasion. Upon M. bovis BCG infection, the nucleomodulin MgdE is delivered into the host nucleus via its NLS and directly binds to the COMPASS complex subunits, ASH2L or WDR5. This interaction disrupts H3K4 trimethylation (H3K4me3) deposition, leading to the epigenetic suppression of pro-inflammatory cytokine transcription (e.g. IL6), thereby facilitating the intracellular survival of the pathogen.
Figure 7—figure supplement 1
MgdE facilitates bacterial colonization in the spleens of infected mice.
Bacterial colonization was assessed in splenic homogenates from infected mice (as described in Figure 7A) by quantifying bacterial DNA using quantitative PCR at 2, 14, 21, 28, and 56 days post-infection. Data are presented as mean ± SD (n=3). Statistical significance was determined using two-way ANOVA, *p<0.05, **p<0.01, and ***p<0.001.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mycobacterium bovis BCG) | Mycobacterium bovis BCG | Chen et al., 2022 | ATCC: Cat#35734 | |
| Strain, strain background (Escherichia coli) | E. coli DH5a | Wang et al., 2025 | ATCC: Cat#25922 | |
| Strain, strain background (Y2HGold Chemically Competent Cell) | Y2HGold | Clontech | Cat#630498 | GAL4-based yeast strain for two-hybrid assays; see ‘Materials and methods: Y2H assay’ for details. |
| Cell line (Homo sapiens) | HEK293T | Cellosaurus | Cat#CRL-3216 RRID:CVCL_0063 | Authenticated and mycoplasma-free; see ‘Materials and methods: Cell culture’ for details. |
| Cell line (H. sapiens) | THP-1 | Cellosaurus | Cat#TIB-202 RRID:CVCL_0006 | Authenticated and mycoplasma-free; see ‘Materials and methods: Cell culture’ for details. |
| Cell line (Mus musculus) | RAW264.7 | Cellosaurus | Cat#TIB-71 RRID:CVCL_C6XG | Authenticated and mycoplasma-free; see ‘Materials and methods: Cell culture’ for details. |
| Cell line (M. musculus) | C57BL/6 | Chang-sheng Bio (Liaoning, China) | RRID:MGI:2159965 | |
| Antibody | Anti-β-actin (Mouse Monoclonal Antibody) | Abbkine | RRID:AB_3740145 | WB 1:10000 |
| Antibody | Anti-Histone H3 (Mouse Monoclonal Antibody) | Abbkine | RRID:AB_3740146 | WB 1:2000 |
| Antibody | Goat anti-rabbit IgG | Abbkine | RRID:AB_2876889 | WB 1:10000 |
| Antibody | Goat anti-mouse IgG | Abbkine | RRID:AB_2737290 | WB 1:1000 |
| Antibody | GAPDH (Mouse Monoclonal Antibody) | Abbkine | RRID:AB_3714704 | WB 1:2000 |
| Antibody | HA tag (Rabbit Polyclonal Antibody) | Proteintech | RRID:AB_11042321 | WB 1:3000 |
| Antibody | GFP tag (Rabbit Polyclonal Antibody) | Proteintech | RRID:AB_11182611 | WB 1:5000 |
| Antibody | Flag tag (Mouse Monoclonal Antibody) | Proteintech | RRID:AB_2918475 | WB 1:10000 |
| Antibody | Anti-H3K4me3 (Rabbit Monoclonal Antibody) | Proteintech | RRID:AB_3740151 | WB 1:5000 |
| Antibody | Ag85B | Zhang et al., 2022 | N/A | WB 1:10000 |
| Antibody | GlpX | Zhang et al., 2022 | N/A | WB 1:10000 |
| Chemical compound, drug | DMEM | Gibco | Cat#11965092 | Cell culture |
| Chemical compound, drug | RPMI 1640 | Gibco | Cat#C11875500BT | Cell culture |
| Chemical compound, drug | Opti-MEMTM | Gibco | Cat#31985070 | Cell culture |
| Chemical compound, drug | FBS | Gibco | Cat#A5256701 | Cell culture |
| Chemical compound, drug | Sodium pyruvate | Gibco | Cat#11360070 | Cell culture |
| Chemical compound, drug | L-glutamine | Gibco | Cat#A2916801 | Cell culture |
| Chemical compound, drug | HEPES | Gibco | Cat#15630080 | Cell culture |
| Chemical compound, drug | 2-Mercaptoethanol | Gibco | Cat#21985023 | Cell culture |
| Chemical compound, drug | Penicillin-streptomycin antibiotics | Gibco | Cat#15140122 | Cell culture |
| Chemical compound, drug | PMA | Sigma-Aldrich | Cat#P8139 | Cell culture |
| Chemical compound, drug | DO Supplement -Leu/-Trp | Coolaber | Cat#PM2220 | Cell culture |
| Chemical compound, drug | DO Supplement -Ade-His-Leu-Trp | Coolaber | Cat#PM2110 | Y2H strains culture |
| Chemical compound, drug | Aureobasidin A | Coolaber | Cat#CA2332G | Y2H strains culture |
| Chemical compound, drug | OADC | MilliporeSigma | Cat#M0678 | Y2H strains culture |
| Chemical compound, drug | NP-40 | Beyotime | Cat#P0038 | Immunoblotting assay |
| Chemical compound, drug | RIPA buffer | Beyotime | Cat#P0038 | Immunoblotting assay |
| Chemical compound, drug | Protease inhibitor cocktail | Boster | Cat#AR1182 | Immunoblotting assay |
| Chemical compound, drug | Triton-X-100 | Beyotime | Cat#P0096 | Immunoblotting assay |
| Commercial assay or kit | Nuclear and Cytoplasmic Protein Extraction kit | Beyotime | Cat#P0027 | Immunoblotting assay |
| Chemical compound, drug | DAPI | Beyotime | Cat#P0126 | confocal microscopy assay |
| Commercial assay or kit | Clarity Western ECL Substrate | BIO-RAD | Cat#1705060 | confocal microscopy assay |
| Chemical compound, drug | Protein A/G magnetic beads | MedChemExpress | Cat#HY-K0202 | Co-IP assay |
| Chemical compound, drug | Protease inhibitor cocktail | Boster | Cat#AR1182 | Co-IP assay |
| Chemical compound, drug | HieffTrans Liposomal Transfection Reagent | YEASEN | Cat#40802ES03 | Cell fractionation |
| Commercial assay or kit | EndoFree Plasmid Mini Kit | Aidlab | Cat#PL0401 | Cell fractionation |
| Commercial assay or kit | TRIpure Reagent | Aidlab | Cat#RN0101 | qRT-PCR assay |
| Commercial assay or kit | EASYspin RNA Mini Kit | Aidlab | Cat#RN0702 | qRT-PCR assay |
| Commercial assay or kit | cDNA Reverse Transcription Kit | Vazyme | Cat#R333-01 | qRT-PCR assay |
| Commercial assay or kit | 2× ChamQ Universal SYBR qPCR Master Mix | Vazyme | Cat#Q711-03 | qRT-PCR assay |
| Software, algorithm | AlphaFold v2.2.0 | SciCrunch Registry | RRID:SCR_023662 | |
| Software, algorithm | GraphPad Prism 8 | SciCrunch Registry | RRID:SCR_002798 | |
| Software, algorithm | UCSF Chimera | SciCrunch Registry | RRID:SCR_004097 | |
| Software, algorithm | MEGA 12.0 | SciCrunch Registry | RRID:SCR_000667 | |
| Software, algorithm | Cytoscape 3.10.3 | SciCrunch Registry | RRID:SCR_003032 | |
| Software, algorithm | CaseViewer v2.0 | SciCrunch Registry | RRID:SCR_017654 | |
| Software, algorithm | jvenn/Venny 2.1.0 | SciCrunch Registry | RRID:SCR_016343 | |
| Software, algorithm | cNLS Mapper | N/A | N/A | |
| Software, algorithm | iTOL | SciCrunch Registry | RRID:SCR_018174 | |
| Software, algorithm | ESPript | SciCrunch Registry | RRID:SCR_006587 | |
| Software, algorithm | SignalP 5.0 | SciCrunch Registry | RRID:SCR_015644 | |
| Software, algorithm | SecretomeP 2.0 | SciCrunch Registry | RRID:SCR_026505 | |
| Software, algorithm | STRING 12.0 | SciCrunch Registry | RRID:SCR_005223 |
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/107677/elife-107677-mdarchecklist1-v1.docx
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Supplementary file 1
Five Mycobacterium species secreted proteins.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp1-v1.xlsx
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Supplementary file 2
Five Mycobacterium species with conserved nucleomodulins.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp2-v1.xlsx
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Supplementary file 3
56 candidate protein nuclear phenotypes.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp3-v1.xlsx
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Supplementary file 4
Primers used in this study.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp4-v1.xlsx
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Supplementary file 5
Plasmids used in this study.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp5-v1.xlsx
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Supplementary file 6
Bacterial strains used in this study.
- https://cdn.elifesciences.org/articles/107677/elife-107677-supp6-v1.xlsx
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A conserved mycobacterial nucleomodulin hijacks the host COMPASS complex to reprogram pro-inflammatory transcription and promote intracellular survival
eLife 14:RP107677.
https://doi.org/10.7554/eLife.107677.4
