Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B
Background: <i>Staphylococcus aureus</i> is a leading cause of contagious mastitis in dairy cattle. Internalization of <i>S. aureus</i> by bovine mammary gland epithelial cells is thought to be responsible for persistent and chronic intramammary infection, but the underlying...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-10-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/9/10/2090 |
| _version_ | 1827679037547347968 |
|---|---|
| author | Yi-Tian Ying Wei-Jia Ren Xun Tan Jing Yang Rui Liu Ai-Fang Du |
| author_facet | Yi-Tian Ying Wei-Jia Ren Xun Tan Jing Yang Rui Liu Ai-Fang Du |
| author_sort | Yi-Tian Ying |
| collection | DOAJ |
| description | Background: <i>Staphylococcus aureus</i> is a leading cause of contagious mastitis in dairy cattle. Internalization of <i>S. aureus</i> by bovine mammary gland epithelial cells is thought to be responsible for persistent and chronic intramammary infection, but the underlying mechanisms are not fully understood. Methods: In the present study, we evaluated the role of Annexin A2 (AnxA2), a membrane-binding protein, in <i>S. aureus</i> invasion into bovine mammary epithelial cell line (MAC-T). In vitro binding assays were performed to co-immunoprecipitate the binding proteins of AnxA2 in the lysates of <i>S. aureus</i>. Results: AnxA2 mediated the internalization but not adherence of <i>S. aureus</i>. Engagement of AnxA2 stimulated an integrin-linked protein kinase (ILK)/p38 MAPK cascade to induce <i>S. aureus</i> invasion. One of the AnxA2-precipitated proteins was identified as <i>S. aureus</i> clumping factor B (ClfB) through use of mass spectrometry. Direct binding of ClfB to AnxA2 was further confirmed by using a pull-down assay. Pre-incubation with recombinant ClfB protein enhanced <i>S. aureus</i> internalization, an effect that was specially blocked by anti-AnxA2 antibody. Conclusion: Our results demonstrate that binding of ClfB to AnxA2 has a function in promoting <i>S. aureus</i> internalization. Targeting the interaction of ClfB and AnxA2 may confer protection against <i>S. aureus</i> mastitis. |
| first_indexed | 2024-03-10T06:22:07Z |
| format | Article |
| id | doaj.art-821b3d7578934edbba544bc5d0c1bbd0 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T06:22:07Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-821b3d7578934edbba544bc5d0c1bbd02023-11-22T19:14:07ZengMDPI AGMicroorganisms2076-26072021-10-01910209010.3390/microorganisms9102090Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor BYi-Tian Ying0Wei-Jia Ren1Xun Tan2Jing Yang3Rui Liu4Ai-Fang Du5Department of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaDepartment of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaDepartment of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaDepartment of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaDepartment of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaDepartment of Veterinary Medicine, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, ChinaBackground: <i>Staphylococcus aureus</i> is a leading cause of contagious mastitis in dairy cattle. Internalization of <i>S. aureus</i> by bovine mammary gland epithelial cells is thought to be responsible for persistent and chronic intramammary infection, but the underlying mechanisms are not fully understood. Methods: In the present study, we evaluated the role of Annexin A2 (AnxA2), a membrane-binding protein, in <i>S. aureus</i> invasion into bovine mammary epithelial cell line (MAC-T). In vitro binding assays were performed to co-immunoprecipitate the binding proteins of AnxA2 in the lysates of <i>S. aureus</i>. Results: AnxA2 mediated the internalization but not adherence of <i>S. aureus</i>. Engagement of AnxA2 stimulated an integrin-linked protein kinase (ILK)/p38 MAPK cascade to induce <i>S. aureus</i> invasion. One of the AnxA2-precipitated proteins was identified as <i>S. aureus</i> clumping factor B (ClfB) through use of mass spectrometry. Direct binding of ClfB to AnxA2 was further confirmed by using a pull-down assay. Pre-incubation with recombinant ClfB protein enhanced <i>S. aureus</i> internalization, an effect that was specially blocked by anti-AnxA2 antibody. Conclusion: Our results demonstrate that binding of ClfB to AnxA2 has a function in promoting <i>S. aureus</i> internalization. Targeting the interaction of ClfB and AnxA2 may confer protection against <i>S. aureus</i> mastitis.https://www.mdpi.com/2076-2607/9/10/2090<i>Staphylococcus aureus</i>annexin A2clumping factor Bmammary epithelial cells |
| spellingShingle | Yi-Tian Ying Wei-Jia Ren Xun Tan Jing Yang Rui Liu Ai-Fang Du Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B Microorganisms <i>Staphylococcus aureus</i> annexin A2 clumping factor B mammary epithelial cells |
| title | Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B |
| title_full | Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B |
| title_fullStr | Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B |
| title_full_unstemmed | Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B |
| title_short | Annexin A2-Mediated Internalization of <i>Staphylococcus aureus</i> into Bovine Mammary Epithelial Cells Requires Its Interaction with Clumping Factor B |
| title_sort | annexin a2 mediated internalization of i staphylococcus aureus i into bovine mammary epithelial cells requires its interaction with clumping factor b |
| topic | <i>Staphylococcus aureus</i> annexin A2 clumping factor B mammary epithelial cells |
| url | https://www.mdpi.com/2076-2607/9/10/2090 |
| work_keys_str_mv | AT yitianying annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb AT weijiaren annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb AT xuntan annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb AT jingyang annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb AT ruiliu annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb AT aifangdu annexina2mediatedinternalizationofistaphylococcusaureusiintobovinemammaryepithelialcellsrequiresitsinteractionwithclumpingfactorb |