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TitleThermophilic bacteria employ a contractile injection system in hot spring microbial mats.
Journal, issue, pagesISME J, Year 2026
Publish dateFeb 10, 2026
AuthorsVasil A Gaisin / Corina Hadjicharalambous / Izabela Mujakić / Cristian Villena-Alemany / Jiangning Li / Michal Koblížek / Martin Pilhofer /
PubMed AbstractBacterial contractile injection systems (CISs) are multiprotein complexes that facilitate the bacterial response to environmental factors or interactions with other organisms. Multiple novel CISs ...Bacterial contractile injection systems (CISs) are multiprotein complexes that facilitate the bacterial response to environmental factors or interactions with other organisms. Multiple novel CISs have been characterised in laboratory bacterial cultures recently; however, studying CISs in the context of the native microbial community remains challenging. Here, we present an approach to characterise a bioinformatically predicted CIS by directly analysing bacterial cells from their natural environment. Using cryo-focused ion beam milling and cryo-electron tomography (cryoET) imaging, guided by 16S rRNA gene amplicon sequencing, we discovered that thermophilic Chloroflexota bacteria produce intracellular CIS particles in a natural hot spring microbial mat. We then found a niche-specific production of CIS in the structured microbial community using an approach combining metagenomics, proteomics, and immunogold staining. Bioinformatic analysis and imaging revealed CISs in other extremophilic Chloroflexota and Deinococcota. This Chloroflexota/Deinococcota CIS lineage shows phylogenetic and structural similarity to previously described cytoplasmic CIS from Streptomyces and probably shares the same cytoplasmic mode of action. Our integrated environmental cryoET approach is suitable for discovering and characterising novel macromolecular complexes in environmental samples.
External linksISME J / PubMed:41665259
MethodsEM (tomography)
Structure data

EMDB-55835: Lysed Roseiflexus cells from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55839: Roseiflexus cells from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55841: Roseiflexus cells from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55842: Roseiflexus cell from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55846: Roseiflexus cell from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55848: Roseiflexus cell from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55851: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55852: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55853: Roseiflexus cell from microbial mat with contracted contractile injection systems.
Method: EM (tomography)

EMDB-55854: Calidithermus chliarophilus cells with contractile injection systems.
Method: EM (tomography)

EMDB-55855: Deinococcus aquatilis cells with contractile injection systems.
Method: EM (tomography)

EMDB-55861: Roseiflexus castenholzii cells with contractile injection systems.
Method: EM (tomography)

EMDB-55862: Roseiflexus castenholzii cells with contractile injection systems.
Method: EM (tomography)

EMDB-55870: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55871: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55872: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

EMDB-55873: Roseiflexus cells from microbial mat with contractile injection systems.
Method: EM (tomography)

Source
  • Roseiflexus (bacteria)
  • Calidithermus chliarophilus DSM 9957 (bacteria)
  • Deinococcus aquatilis (bacteria)
  • Roseiflexus castenholzii DSM 13941 (bacteria)

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