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Reposted by Jordan Jastrab

Ryan Flynn @raflynn5.bsky.social · 1d

We’re excited to report work led by postdoc Jennifer Porat in the lab, finding that DNA accumulates on the surface of living cells and that the secreted extracellular protein DNASE1L3 can modulate its levels on B and T cells. With a new twist for ATAC-seq as well www.biorxiv.org/content/10.1...

DNASE1L3 surveils mitochondrial DNA on the surface of distinct mammalian cells

The extracellular space is a critical environment for discriminating self versus non-self nucleic acids and initiating the appropriate immune responses through signaling cascades to relay information ...

www.biorxiv.org

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

A huge thanks to co-authors @ashleytseng.bsky.social , @danielhfisch.bsky.social, @StephanieRaglandNotOnBlueSky, fearless leader @jkagan1.bsky.social, and to colleagues who gave advice and strains so I could learn to work with Staph. Feel free to comment or DM with any questions/suggestions!

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

WTA here, despite being “immunogenic” in its ability to promote immune responses, does not directly activate immune receptors. Rather, it regulates availability of the PAMP DNA, essentially acting as a “meta-PAMP” akin to Legionella meta-effectors that regulate activity of direct-acting effectors.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Collectively these data suggest that in contrast to toxigenic S. aureus infection where NLRP3 is robustly activated, during nontoxigenic infection S. aureus protects itself from being detected by cytosolic DNA sensors by O-acetylating its PGN, thus limiting WTA abundance on its cell surface.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Finally, we tested if differences in bacterial DNA release impact other immune DNA sensors as well, in particular cGAS. Using cells designed and generated by my incredible colleague Stephanie Ragland, we found that OatA and TarMS affect cGAS activation in the same way they affect AIM2 activity.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Our finding that S. aureus activates a DNA sensor suggested OatA and TarMS regulate DNA release within infected cells. We purified and quantified extrabacterial DNA from infected macrophages, and consistent with our hypothesis found oatA mutants released more whereas tarMS mutants released less DNA.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Using an AIM2-mScarlet fluorescent fusion protein and GFP-producing bacteria, incredible PhD student @ashleytseng.bsky.social with a major assist from @danielhfisch.bsky.social found that AIM2 specks (red), representing inflammasomes, form in close proximity to bacteria (green).

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

To clarify the mechanism by which WTA enhances inflammasome activation we turned to the host, reasoning that identifying the receptor required would lead us to the ligand that WTA controls. We identified DNA sensor AIM2 as the receptor activated during nontoxigenic S. aureus infection.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

We then returned to TarM, which glycosylates WTA. Glycosylation of WTA is increasingly appreciated to play important roles in host-microbe interactions. We knocked out glycosyltransferases TarM and TarS and found that both promote inflammasome activation, with TarM having a more potent effect.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Further supporting our hypothesis, chemically blocking WTA synthesis ablated inflammasome activation and eliminated the difference between WT and oatA strains. Thus, WTA is essential for inflammasome activation by nontoxigenic S. aureus, and is required for the immunoevasive function of OatA.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

TarM is an enzyme that modifies cell wall component wall teichoic acid (WTA). Strikingly, OatA acetylates the identical PGN site that WTA is attached to. We found that oatA mutants have more WTA than WT bacteria, suggesting the way OatA reduces inflammasome activation is by reducing WTA abundance.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

We hypothesized OatA instead restricts the activity of a bacterial factor that promotes inflammasome activation. To identify such a factor, we generated a transposon mutant library in the oatA mutant background and isolated mutants defective for activating inflammasomes; we identified a tarM mutant.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

PGN acetylation by OatA blocks bacterial killing by phagosomal lysozyme; this was thought to be how OatA prevents inflammasome activation. But we knocked out lysozyme from iBMDMs and found it was dispensable for inflammasome activity. OatA’s role here is thus distinct from its established function.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Prior studies have shown bacteria lacking PGN O-acetyltransferase OatA elicit greater inflammasome activation than WT bacteria. We rederived an oatA mutant in a nontoxigenic S. aureus background and reproduced the finding that OatA helps S. aureus evade inflammasomes during infection of macrophages.

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

S. aureus secretes toxins that kill host cells and activate the NLRP3 inflammasome. After Staph spreads through the blood it reduces toxin production and can survive inside host cells, triggering minimal immunity. How does nontoxigenic Staph avoid activating inflammasomes within infected cells?

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Now that I am no longer dealing with an infant with an infection (non-staphylococcal), time for a proper 🧵! If you’re interested in some combination of Staphylococcus aureus, innate immune sensing of bacteria, inflammasomes, and cell wall modifications, read on…

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Jordan Jastrab @jordanjastrab.bsky.social · 10d

Thanks so much Brian! I hope you and Irma are both doing well yourselves!!

Reposted by Jordan Jastrab

Jonathan Kagan @jkagan1.bsky.social · 12d

Notably, WTA is not a PAMP.
Rather, WTA controls the release of bacterial DNA into cells, where it can act as a PAMP for AIM2 and cGAS. How many other meta-PAMPs are out there?
@jordanjastrab.bsky.social @harvardmed.bsky.social @bostonchildrens.bsky.social @harvardmicro.bsky.social

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Jordan Jastrab @jordanjastrab.bsky.social · 12d

☺️☺️☺️

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Jordan Jastrab @jordanjastrab.bsky.social · 12d

Thanks Lauren!!

Jordan Jastrab @jordanjastrab.bsky.social · 12d

Thrilled to share my work from the @jkagan1.bsky.social lab!

TLDR: O-acetylation and wall teichoic acid compete for space on S. aureus peptidoglycan. This competition regulates DNA release, and thus immune receptor activation, during infection!

www.biorxiv.org/content/10.1...

#microsky #immunosky

Wall teichoic acid is required for DNA-triggered innate immune receptor activation by Staphylococcus aureus

Receptors that stimulate inflammation are commonly activated by ligands that are buried within microbial cells. The mechanisms that facilitate immunostimulatory ligand release from microbes during inf...

www.biorxiv.org

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Reposted by Jordan Jastrab

Boston Bacterial Meeting @bostonbacteria.bsky.social · Feb 1

SAVE THE DATE ‼️ 📆 📢
BBM 2025 will be held on June 9-10th at the Harvard Science Center feat. the one and only Dr. Petra Levin as keynote! We can't wait to see you there.

Registration and scholarship applications open next week.

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Reposted by Jordan Jastrab

Vijay Rathinam @vijayrathinam.bsky.social · Dec 31

So excited to have our new paper out now!
Pyroptotic EVs “transplant” gasdermin pores from dying cells onto bystander cells, propagating pyroptosis and inflammation. t.co/XgU6jt15aF 1/3
#innate_immunity #immunoSky #celldeath #ExtracellularVesicles @uconnresearch.bsky.social

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Reposted by Jordan Jastrab

Martin Hunt @martibartfast.bsky.social · Nov 29

I just made a first release of TNA - new tool to compare 2 bacterial genomes. Is shamelessly inspired by ACT, but should be simpler to use. Drag-n-drop two genomes: it runs BLAST for you and then you can visualize. For Mac, Windows 11, Linux tna.readthedocs.io

TNA documentation — TNA 0.0.1 documentation

tna.readthedocs.io

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Reposted by Jordan Jastrab

ᐯIᑕTOᖇ ᑎIᘔET, ᗰᗪ @nizet.bsky.social · Nov 24

Wow—one of the most impressive bench-to-bedside stories you’re ever going to see out today @Nature

Spatial proteomics of potentially fatal toxic epidermal necrolysis/Stevens-Johnson Syndrome drug reactions identifies JAK/STAT pathway inhibition as effective therapy!

www.nature.com/articles/s41...

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