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| Status |
Public on Mar 27, 2023 |
| Title |
Bacteriophage antidefense genes that neutralize TIR and STING immune responses |
| Organism |
Bacteriophage sp. |
| Experiment type |
Other
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| Summary |
Programmed cell suicide of infected bacteria, known as abortive infection (Abi), serves as a central immune defense strategy to prevent the spread of bacteriophage viruses and other invasive genetic elements across a population. Many Abi systems utilize bespoke cyclic nucleotide immune messengers generated upon infection to rapidly mobilize cognate death effectors. Here, we identify a large family of bacteriophage nucleotidyltransferases (NTases) which synthesize competitor cyclic dinucleotide (CDN) ligands and inhibit NAD-depleting TIR effectors activated through a linked STING CDN sensor domain (TIR-STING). Through a functional screen of NTase-adjacent phage genes, we uncover candidate inhibitors of host TIR-STING suicide signaling. Among these, we demonstrate that a virus MazG-like nucleotide pyrophosphatase, Atd1, depletes the starvation alarmone (p)ppGpp, revealing a role for the alarmone-activated host toxin MazF as a key executioner of TIR-driven abortive infection. Phage NTases and counter-defenses like Atd1 preserve host viability to ensure virus propagation, and may be exploited as tools to modulate TIR and STING immune responses.
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| Overall design |
The pooled phage ORF library was transformed into BL21 (DE3) Rosetta 2 competent cells (MilliporeSigma) together with pET30a encoding Flag-TIR-STING or empty vector (EV) control.
100 ml of overnight cultures were diluted 1:20 and grown in M9 media supplemented with 0.25% CAA and 1% glycerol, and induced at A600=0.3-0.4 through the addition of 0.5mM IPTG. Samples were taken immediately prior to induction and at 18 h post-induction.
Reads were obtained from DNA isolated from cultures containing EV or TIR-STING, at either time 0 or after 18 hr induction.
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| Web link |
https://pubmed.ncbi.nlm.nih.gov/36952342/
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| Contributor(s) |
Ho P, Chen Y, Biswas S, Canfield E, Abdolvahabi A, Feldman DE |
| Citation(s) |
36952342 |
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| Submission date |
Jan 03, 2023 |
| Last update date |
Jun 26, 2023 |
| Contact name |
Douglas Edmund Feldman |
| E-mail(s) |
defeldma@usc.edu
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| Organization name |
USC Keck School of Medicine
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| Department |
Pathology
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| Lab |
HMR 212
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| Street address |
2011 Zonal Avenue
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| City |
Los Angeles |
| State/province |
CA |
| ZIP/Postal code |
90033 |
| Country |
USA |
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| Platforms (1) |
| GPL32997 |
Illumina HiSeq 3000 (Bacteriophage sp.) |
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| Samples (6)
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| GSM6914211 |
TIR-STING, baseline- 0 hr |
| GSM6914212 |
TIR-STING induced, harvested at 18 hr, replicate 1 |
| GSM6914213 |
TIR-STING induced, harvested at 18 hr, replicate 2 |
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| Relations |
| BioProject |
PRJNA917585 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE222071_Partek_ampliconseq_Aug2021_Normalization_Normalized_counts_CPM_v5.txt.gz |
5.7 Kb |
(ftp)(http) |
TXT |
| GSE222071_amplicon_sequence.fa.gz |
20.4 Kb |
(ftp)(http) |
FA |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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