Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems

Abstract CRISPR-Cas and restriction-modification systems represent the core defense arsenal in Streptococcus thermophilus, but their effectiveness is compromised by phages encoding anti-CRISPR proteins (ACRs) and other counter-defense strategies. Here, we explore the defensome of 263 S. thermophilus...

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Main Authors: Audrey Leprince, Justine Lefrançois, Anne M. Millen, Damian Magill, Philippe Horvath, Dennis A. Romero, Geneviève M. Rousseau, Sylvain Moineau
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62408-3
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author Audrey Leprince
Justine Lefrançois
Anne M. Millen
Damian Magill
Philippe Horvath
Dennis A. Romero
Geneviève M. Rousseau
Sylvain Moineau
author_facet Audrey Leprince
Justine Lefrançois
Anne M. Millen
Damian Magill
Philippe Horvath
Dennis A. Romero
Geneviève M. Rousseau
Sylvain Moineau
author_sort Audrey Leprince
collection DOAJ
description Abstract CRISPR-Cas and restriction-modification systems represent the core defense arsenal in Streptococcus thermophilus, but their effectiveness is compromised by phages encoding anti-CRISPR proteins (ACRs) and other counter-defense strategies. Here, we explore the defensome of 263 S. thermophilus strains to uncover other anti-phage systems. The defense landscape of S. thermophilus is enriched by 21 accessory defense systems, 13 of which have never been investigated in this species. Experimental validation of 17 systems with 14 phages reveals a range of anti-phage activities, highlighting both broad and narrow specificities across the five viral genera infecting S. thermophilus. Synergies are observed when combining CRISPR immunity with accessory systems. We also assess the fitness cost associated with the chromosomal integration of these systems in their native context and find no impact under laboratory or industrial conditions. These findings underscore the potential of these accessory defense systems to enhance the resistance of S. thermophilus, particularly against ACR-encoding phages.
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series Nature Communications
spelling doaj-art-cfe95623a86f4f35b87fb745c4d782ed2025-08-20T03:46:15ZengNature PortfolioNature Communications2041-17232025-08-0116111610.1038/s41467-025-62408-3Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systemsAudrey Leprince0Justine Lefrançois1Anne M. Millen2Damian Magill3Philippe Horvath4Dennis A. Romero5Geneviève M. Rousseau6Sylvain Moineau7Département de biochimie, de microbiologie et de bio-informatique, Faculté des sciences et de génie, Université Laval, Québec City, QC, Canada. Institut de Biologie Intégrative et des Systèmes (IBIS), Pavillon Charles-Eugène-Marchand, Université LavalDépartement de biochimie, de microbiologie et de bio-informatique, Faculté des sciences et de génie, Université Laval, Québec City, QC, Canada. Institut de Biologie Intégrative et des Systèmes (IBIS), Pavillon Charles-Eugène-Marchand, Université LavalIFF Health and Biosciences, 3329 Agriculture DrIFF Health and Biosciences, CS 10010IFF Health and Biosciences, CS 10010IFF Health and Biosciences, 3329 Agriculture DrDépartement de biochimie, de microbiologie et de bio-informatique, Faculté des sciences et de génie, Université Laval, Québec City, QC, Canada. Institut de Biologie Intégrative et des Systèmes (IBIS), Pavillon Charles-Eugène-Marchand, Université LavalDépartement de biochimie, de microbiologie et de bio-informatique, Faculté des sciences et de génie, Université Laval, Québec City, QC, Canada. Institut de Biologie Intégrative et des Systèmes (IBIS), Pavillon Charles-Eugène-Marchand, Université LavalAbstract CRISPR-Cas and restriction-modification systems represent the core defense arsenal in Streptococcus thermophilus, but their effectiveness is compromised by phages encoding anti-CRISPR proteins (ACRs) and other counter-defense strategies. Here, we explore the defensome of 263 S. thermophilus strains to uncover other anti-phage systems. The defense landscape of S. thermophilus is enriched by 21 accessory defense systems, 13 of which have never been investigated in this species. Experimental validation of 17 systems with 14 phages reveals a range of anti-phage activities, highlighting both broad and narrow specificities across the five viral genera infecting S. thermophilus. Synergies are observed when combining CRISPR immunity with accessory systems. We also assess the fitness cost associated with the chromosomal integration of these systems in their native context and find no impact under laboratory or industrial conditions. These findings underscore the potential of these accessory defense systems to enhance the resistance of S. thermophilus, particularly against ACR-encoding phages.https://doi.org/10.1038/s41467-025-62408-3
spellingShingle Audrey Leprince
Justine Lefrançois
Anne M. Millen
Damian Magill
Philippe Horvath
Dennis A. Romero
Geneviève M. Rousseau
Sylvain Moineau
Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
Nature Communications
title Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
title_full Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
title_fullStr Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
title_full_unstemmed Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
title_short Strengthening phage resistance of Streptococcus thermophilus by leveraging complementary defense systems
title_sort strengthening phage resistance of streptococcus thermophilus by leveraging complementary defense systems
url https://doi.org/10.1038/s41467-025-62408-3
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