Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells
ABSTRACTOuter membrane vesicles (OMVs) are proteoliposomes shed by Gram-negative bacteria. Their secretion is enhanced by the transition into the intra-host milieu, and OMVs play critical roles during pathogenesis. Enterohemorrhagic Escherichia coli O157 (EHEC) can cause diarrheal disease in humans,...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-12-01
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| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msphere.00520-23 |
| _version_ | 1797384694373810176 |
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| author | Natalie Sirisaengtaksin Eloise J. O'Donoghue Sara Jabbari Andrew J. Roe Anne Marie Krachler |
| author_facet | Natalie Sirisaengtaksin Eloise J. O'Donoghue Sara Jabbari Andrew J. Roe Anne Marie Krachler |
| author_sort | Natalie Sirisaengtaksin |
| collection | DOAJ |
| description | ABSTRACTOuter membrane vesicles (OMVs) are proteoliposomes shed by Gram-negative bacteria. Their secretion is enhanced by the transition into the intra-host milieu, and OMVs play critical roles during pathogenesis. Enterohemorrhagic Escherichia coli O157 (EHEC) can cause diarrheal disease in humans, and soluble toxins including Shiga-like toxins that contribute to disease severity and clinical complications like hemolytic uremic syndrome have been shown to be OMV associated. In addition to Shiga-like toxins, EHEC produces a type III secretion system (T3SS), and T3SS effectors are associated with colonization and disease severity in vivo. Here, we show that type III secreted substrates including translocators and effectors are incorporated into OMVs both in the presence and absence of a functional T3SS. EHEC strains with non-functional T3SS shed more OMVs, and vesicles enter host cells with accelerated kinetics compared to vesicles shed from wild-type EHEC. The T3SS effector translocated intimin receptor (Tir) is trafficked from OMVs into host cells and localizes to the membrane. However, its clustering on the host membrane and co-localization with bacterial pedestals is intimin dependent. We show that OMV-delivered Tir can cross-complement an effector-deficient EHEC strain and promote pedestal formation. Together, these data demonstrate that OMVs provide an alternative pathway for the delivery of EHEC T3SS cargo and that vesicle associated effectors are released from OMVs inside host cells and can retain biological activity.IMPORTANCEBacteria can package protein cargo into nanosized membrane blebs that are shed from the bacterial membrane and released into the environment. Here, we report that a type of pathogenic bacteria called enterohemorrhagic Escherichia coli O157 (EHEC) uses their membrane blebs (outer membrane vesicles) to package components of their type 3 secretion system and send them into host cells, where they can manipulate host signaling pathways including those involved in infection response, such as immunity. Usually, EHEC use a needle-like apparatus to inject these components into host cells, but packaging them into membrane blebs that get taken up by host cells is another way of delivery that can bypass the need for a functioning injection system. |
| first_indexed | 2024-03-08T21:40:22Z |
| format | Article |
| id | doaj.art-b235a442b5504fafa8012d2a6503453f |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-03-08T21:40:22Z |
| publishDate | 2023-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-b235a442b5504fafa8012d2a6503453f2023-12-20T14:01:03ZengAmerican Society for MicrobiologymSphere2379-50422023-12-018610.1128/msphere.00520-23Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cellsNatalie Sirisaengtaksin0Eloise J. O'Donoghue1Sara Jabbari2Andrew J. Roe3Anne Marie Krachler4Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas, USASchool of Biosciences, Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, United KingdomSchool of Mathematics, Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, United KingdomSchool of Infection and Immunity, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United KingdomDepartment of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas, USAABSTRACTOuter membrane vesicles (OMVs) are proteoliposomes shed by Gram-negative bacteria. Their secretion is enhanced by the transition into the intra-host milieu, and OMVs play critical roles during pathogenesis. Enterohemorrhagic Escherichia coli O157 (EHEC) can cause diarrheal disease in humans, and soluble toxins including Shiga-like toxins that contribute to disease severity and clinical complications like hemolytic uremic syndrome have been shown to be OMV associated. In addition to Shiga-like toxins, EHEC produces a type III secretion system (T3SS), and T3SS effectors are associated with colonization and disease severity in vivo. Here, we show that type III secreted substrates including translocators and effectors are incorporated into OMVs both in the presence and absence of a functional T3SS. EHEC strains with non-functional T3SS shed more OMVs, and vesicles enter host cells with accelerated kinetics compared to vesicles shed from wild-type EHEC. The T3SS effector translocated intimin receptor (Tir) is trafficked from OMVs into host cells and localizes to the membrane. However, its clustering on the host membrane and co-localization with bacterial pedestals is intimin dependent. We show that OMV-delivered Tir can cross-complement an effector-deficient EHEC strain and promote pedestal formation. Together, these data demonstrate that OMVs provide an alternative pathway for the delivery of EHEC T3SS cargo and that vesicle associated effectors are released from OMVs inside host cells and can retain biological activity.IMPORTANCEBacteria can package protein cargo into nanosized membrane blebs that are shed from the bacterial membrane and released into the environment. Here, we report that a type of pathogenic bacteria called enterohemorrhagic Escherichia coli O157 (EHEC) uses their membrane blebs (outer membrane vesicles) to package components of their type 3 secretion system and send them into host cells, where they can manipulate host signaling pathways including those involved in infection response, such as immunity. Usually, EHEC use a needle-like apparatus to inject these components into host cells, but packaging them into membrane blebs that get taken up by host cells is another way of delivery that can bypass the need for a functioning injection system.https://journals.asm.org/doi/10.1128/msphere.00520-23outer membrane vesiclesEHECOMVvesicular traffickingsecretion systems |
| spellingShingle | Natalie Sirisaengtaksin Eloise J. O'Donoghue Sara Jabbari Andrew J. Roe Anne Marie Krachler Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells mSphere outer membrane vesicles EHEC OMV vesicular trafficking secretion systems |
| title | Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells |
| title_full | Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells |
| title_fullStr | Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells |
| title_full_unstemmed | Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells |
| title_short | Bacterial outer membrane vesicles provide an alternative pathway for trafficking of Escherichia coli O157 type III secreted effectors to epithelial cells |
| title_sort | bacterial outer membrane vesicles provide an alternative pathway for trafficking of escherichia coli o157 type iii secreted effectors to epithelial cells |
| topic | outer membrane vesicles EHEC OMV vesicular trafficking secretion systems |
| url | https://journals.asm.org/doi/10.1128/msphere.00520-23 |
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