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a. AlphaFold predictions of RESIST with <t>CNOT1</t> M-HEAT and CNOT9. TTP peptide:CNOT1 M-HEAT is PDB code 4J8S . Roquin peptide:CNOT9 is PDB code 5LSW . N and C-terminal ends of RESIST are marked by “N” or “C” respectively. b. Ifnb1 RT-qPCR from BMMs electroporated with non-targeting control gRNA (NTC) or gRNAs targeting indicated CCR4-NOT subunits, treated for 8 hours with 100 μg/mL DMXAA. c. Ifnb1 RT-qPCR for BMMs lentivirally transduced with indicated constructs, treated with doxycycline and DMXAA for 6 hours. Results include data also shown in . d. Immunoblot for anti-HA IP of BMMs transduced with indicated constructs in e. RESIST construct is C-terminally tagged with HA. e. Ifnb1 RT-qPCR from BMMs electroporated with indicated gRNAs, treated for 8 hours with 100 μg/mL DMXAA.
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a. AlphaFold predictions of RESIST with <t>CNOT1</t> M-HEAT and CNOT9. TTP peptide:CNOT1 M-HEAT is PDB code 4J8S . Roquin peptide:CNOT9 is PDB code 5LSW . N and C-terminal ends of RESIST are marked by “N” or “C” respectively. b. Ifnb1 RT-qPCR from BMMs electroporated with non-targeting control gRNA (NTC) or gRNAs targeting indicated CCR4-NOT subunits, treated for 8 hours with 100 μg/mL DMXAA. c. Ifnb1 RT-qPCR for BMMs lentivirally transduced with indicated constructs, treated with doxycycline and DMXAA for 6 hours. Results include data also shown in . d. Immunoblot for anti-HA IP of BMMs transduced with indicated constructs in e. RESIST construct is C-terminally tagged with HA. e. Ifnb1 RT-qPCR from BMMs electroporated with indicated gRNAs, treated for 8 hours with 100 μg/mL DMXAA.
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Image Search Results


a. AlphaFold predictions of RESIST with CNOT1 M-HEAT and CNOT9. TTP peptide:CNOT1 M-HEAT is PDB code 4J8S . Roquin peptide:CNOT9 is PDB code 5LSW . N and C-terminal ends of RESIST are marked by “N” or “C” respectively. b. Ifnb1 RT-qPCR from BMMs electroporated with non-targeting control gRNA (NTC) or gRNAs targeting indicated CCR4-NOT subunits, treated for 8 hours with 100 μg/mL DMXAA. c. Ifnb1 RT-qPCR for BMMs lentivirally transduced with indicated constructs, treated with doxycycline and DMXAA for 6 hours. Results include data also shown in . d. Immunoblot for anti-HA IP of BMMs transduced with indicated constructs in e. RESIST construct is C-terminally tagged with HA. e. Ifnb1 RT-qPCR from BMMs electroporated with indicated gRNAs, treated for 8 hours with 100 μg/mL DMXAA.

Journal: bioRxiv

Article Title: The SP140-RESIST pathway regulates interferon mRNA stability and antiviral immunity

doi: 10.1101/2024.08.28.610186

Figure Lengend Snippet: a. AlphaFold predictions of RESIST with CNOT1 M-HEAT and CNOT9. TTP peptide:CNOT1 M-HEAT is PDB code 4J8S . Roquin peptide:CNOT9 is PDB code 5LSW . N and C-terminal ends of RESIST are marked by “N” or “C” respectively. b. Ifnb1 RT-qPCR from BMMs electroporated with non-targeting control gRNA (NTC) or gRNAs targeting indicated CCR4-NOT subunits, treated for 8 hours with 100 μg/mL DMXAA. c. Ifnb1 RT-qPCR for BMMs lentivirally transduced with indicated constructs, treated with doxycycline and DMXAA for 6 hours. Results include data also shown in . d. Immunoblot for anti-HA IP of BMMs transduced with indicated constructs in e. RESIST construct is C-terminally tagged with HA. e. Ifnb1 RT-qPCR from BMMs electroporated with indicated gRNAs, treated for 8 hours with 100 μg/mL DMXAA.

Article Snippet: Antibodies used in this study were rat anti-HA (Roche, clone 3F10, 1186742300, 1:1000), mouse anti-actin (Santa Cruz Biotechnology, sc-4478, 1:1000), rabbit anti-CNOT1 (Cell Signaling Technologies, 44613S, 1:1000), rabbit anti-CNOT9 (Proteintech, 22503-1-AP, 1:500), rabbit anti-TTP (Millipore Sigma, ABE285, 1:1000), rabbit anti-CNOT11 (Sigma Aldrich, HPA069823, 0.4 μg/mL), rabbit anti-ZFP36L1 (Cell Signaling Technologies, 30894S, 1:1000), rabbit anti-ZFP36L2 (Abcam, ab70775, 1:1000), and rabbit-SP140 (Covance, as previously described ; 1:1000).

Techniques: Quantitative RT-PCR, Control, Transduction, Construct, Western Blot

a. Schematic of CCR4-NOT subunit and RESIST complexes produced by Alphafold Multimer. b. Aligned AlphaFold models of RESIST with the CNOT1 M-HEAT domain. c. The top-scoring structural prediction of the RESIST and CNOT1 M-HEAT complex, colored by pLDDT. d. AlphaFold PAE plot of predicted RESIST and CNOT1 M-HEAT complex. Plot generated with PAEViewer . e. Depiction of RESIST binding to a hydrophobic patch on the CNOT-1 M-HEAT domain (CNOT1 colored by hydrophobicity).

Journal: bioRxiv

Article Title: The SP140-RESIST pathway regulates interferon mRNA stability and antiviral immunity

doi: 10.1101/2024.08.28.610186

Figure Lengend Snippet: a. Schematic of CCR4-NOT subunit and RESIST complexes produced by Alphafold Multimer. b. Aligned AlphaFold models of RESIST with the CNOT1 M-HEAT domain. c. The top-scoring structural prediction of the RESIST and CNOT1 M-HEAT complex, colored by pLDDT. d. AlphaFold PAE plot of predicted RESIST and CNOT1 M-HEAT complex. Plot generated with PAEViewer . e. Depiction of RESIST binding to a hydrophobic patch on the CNOT-1 M-HEAT domain (CNOT1 colored by hydrophobicity).

Article Snippet: Antibodies used in this study were rat anti-HA (Roche, clone 3F10, 1186742300, 1:1000), mouse anti-actin (Santa Cruz Biotechnology, sc-4478, 1:1000), rabbit anti-CNOT1 (Cell Signaling Technologies, 44613S, 1:1000), rabbit anti-CNOT9 (Proteintech, 22503-1-AP, 1:500), rabbit anti-TTP (Millipore Sigma, ABE285, 1:1000), rabbit anti-CNOT11 (Sigma Aldrich, HPA069823, 0.4 μg/mL), rabbit anti-ZFP36L1 (Cell Signaling Technologies, 30894S, 1:1000), rabbit anti-ZFP36L2 (Abcam, ab70775, 1:1000), and rabbit-SP140 (Covance, as previously described ; 1:1000).

Techniques: Produced, Generated, Binding Assay

a. Aligned AlphaFold structure predictions of RESIST with the CNOT1 NMIF4G-NHEAT domains, CNOT10, and CNOT11. b. Highest-scoring AlphaFold structure prediction of RESIST and CNOT1/CNOT10/CNOT11 complex colored by pLDDT. c. AlphaFold PAE plot of predicted RESIST and CNOT1/CNOT10/CNOT11 complex. Plot generated with PAEViewer .

Journal: bioRxiv

Article Title: The SP140-RESIST pathway regulates interferon mRNA stability and antiviral immunity

doi: 10.1101/2024.08.28.610186

Figure Lengend Snippet: a. Aligned AlphaFold structure predictions of RESIST with the CNOT1 NMIF4G-NHEAT domains, CNOT10, and CNOT11. b. Highest-scoring AlphaFold structure prediction of RESIST and CNOT1/CNOT10/CNOT11 complex colored by pLDDT. c. AlphaFold PAE plot of predicted RESIST and CNOT1/CNOT10/CNOT11 complex. Plot generated with PAEViewer .

Article Snippet: Antibodies used in this study were rat anti-HA (Roche, clone 3F10, 1186742300, 1:1000), mouse anti-actin (Santa Cruz Biotechnology, sc-4478, 1:1000), rabbit anti-CNOT1 (Cell Signaling Technologies, 44613S, 1:1000), rabbit anti-CNOT9 (Proteintech, 22503-1-AP, 1:500), rabbit anti-TTP (Millipore Sigma, ABE285, 1:1000), rabbit anti-CNOT11 (Sigma Aldrich, HPA069823, 0.4 μg/mL), rabbit anti-ZFP36L1 (Cell Signaling Technologies, 30894S, 1:1000), rabbit anti-ZFP36L2 (Abcam, ab70775, 1:1000), and rabbit-SP140 (Covance, as previously described ; 1:1000).

Techniques: Generated

Schematic for how RESIST may interact with CCR4-NOT subunits CNOT1 and CNOT9 to mediate stabilization of Ifnb1 mRNAs.

Journal: bioRxiv

Article Title: The SP140-RESIST pathway regulates interferon mRNA stability and antiviral immunity

doi: 10.1101/2024.08.28.610186

Figure Lengend Snippet: Schematic for how RESIST may interact with CCR4-NOT subunits CNOT1 and CNOT9 to mediate stabilization of Ifnb1 mRNAs.

Article Snippet: Antibodies used in this study were rat anti-HA (Roche, clone 3F10, 1186742300, 1:1000), mouse anti-actin (Santa Cruz Biotechnology, sc-4478, 1:1000), rabbit anti-CNOT1 (Cell Signaling Technologies, 44613S, 1:1000), rabbit anti-CNOT9 (Proteintech, 22503-1-AP, 1:500), rabbit anti-TTP (Millipore Sigma, ABE285, 1:1000), rabbit anti-CNOT11 (Sigma Aldrich, HPA069823, 0.4 μg/mL), rabbit anti-ZFP36L1 (Cell Signaling Technologies, 30894S, 1:1000), rabbit anti-ZFP36L2 (Abcam, ab70775, 1:1000), and rabbit-SP140 (Covance, as previously described ; 1:1000).

Techniques: