Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection
Cytomegalovirus (CMV) infection initiates massive rearrangement of cytoplasmic organelles to generate assembly compartment (AC). The earliest events, the establishment of the preAC, are initiated in the early phase as an extensive reorganization of early endosomes (EEs), endosomal recycling compartm...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-08-01
|
| Series: | Life |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2075-1729/11/9/876 |
| _version_ | 1797518492236251136 |
|---|---|
| author | Igor Štimac Natalia Jug Vučko Gordana Blagojević Zagorac Marina Marcelić Hana Mahmutefendić Lučin Pero Lučin |
| author_facet | Igor Štimac Natalia Jug Vučko Gordana Blagojević Zagorac Marina Marcelić Hana Mahmutefendić Lučin Pero Lučin |
| author_sort | Igor Štimac |
| collection | DOAJ |
| description | Cytomegalovirus (CMV) infection initiates massive rearrangement of cytoplasmic organelles to generate assembly compartment (AC). The earliest events, the establishment of the preAC, are initiated in the early phase as an extensive reorganization of early endosomes (EEs), endosomal recycling compartment (ERC), trans-Golgi network (TGN), and the Golgi. Here, we demonstrate that dynamin inhibitors (Dynasore, Dyngo-4a, MiTMAB, and Dynole-34-2) block the establishment of the preAC in murine CMV (MCMV) infected cells. In this study, we extensively analyzed the effect of Dynasore on the Golgi reorganization sequence into the outer preAC. We also monitored the development of the inner preAC using a set of markers that define EEs (Rab5, Vps34, EEA1, and Hrs), the EE-ERC interface (Rab10), the ERC (Rab11, Arf6), three layers of the Golgi (GRASP65, GM130, Golgin97), and late endosomes (Lamp1). Dynasore inhibited the pericentriolar accumulation of all markers that display EE-ERC-TGN interface in the inner preAC and prevented Golgi unlinking and dislocation to the outer preAC. Furthermore, in pulse-chase experiments, we demonstrated that the presence of dynasore only during the early phase of MCMV infection (4–14 hpi) is sufficient to prevent not only AC formation but also the synthesis of late-phase proteins and virion production. Therefore, our results indicate that dynamin-2 acts as a part of the machinery required for AC generation and rearrangement of EE/ERC/Golgi membranes in the early phase of CMV infection. |
| first_indexed | 2024-03-10T07:30:43Z |
| format | Article |
| id | doaj.art-37d4b8afa340406d9313da287672ab48 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-10T07:30:43Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Life |
| spelling | doaj.art-37d4b8afa340406d9313da287672ab482023-11-22T13:55:14ZengMDPI AGLife2075-17292021-08-0111987610.3390/life11090876Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of InfectionIgor Štimac0Natalia Jug Vučko1Gordana Blagojević Zagorac2Marina Marcelić3Hana Mahmutefendić Lučin4Pero Lučin5Department of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaDepartment of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaDepartment of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaDepartment of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaDepartment of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaDepartment of Physiology and Immunology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, CroatiaCytomegalovirus (CMV) infection initiates massive rearrangement of cytoplasmic organelles to generate assembly compartment (AC). The earliest events, the establishment of the preAC, are initiated in the early phase as an extensive reorganization of early endosomes (EEs), endosomal recycling compartment (ERC), trans-Golgi network (TGN), and the Golgi. Here, we demonstrate that dynamin inhibitors (Dynasore, Dyngo-4a, MiTMAB, and Dynole-34-2) block the establishment of the preAC in murine CMV (MCMV) infected cells. In this study, we extensively analyzed the effect of Dynasore on the Golgi reorganization sequence into the outer preAC. We also monitored the development of the inner preAC using a set of markers that define EEs (Rab5, Vps34, EEA1, and Hrs), the EE-ERC interface (Rab10), the ERC (Rab11, Arf6), three layers of the Golgi (GRASP65, GM130, Golgin97), and late endosomes (Lamp1). Dynasore inhibited the pericentriolar accumulation of all markers that display EE-ERC-TGN interface in the inner preAC and prevented Golgi unlinking and dislocation to the outer preAC. Furthermore, in pulse-chase experiments, we demonstrated that the presence of dynasore only during the early phase of MCMV infection (4–14 hpi) is sufficient to prevent not only AC formation but also the synthesis of late-phase proteins and virion production. Therefore, our results indicate that dynamin-2 acts as a part of the machinery required for AC generation and rearrangement of EE/ERC/Golgi membranes in the early phase of CMV infection.https://www.mdpi.com/2075-1729/11/9/876cytomegalovirusassembly compartmentdynaminearly endosomesendosomal recycling compartmentGolgi apparatus |
| spellingShingle | Igor Štimac Natalia Jug Vučko Gordana Blagojević Zagorac Marina Marcelić Hana Mahmutefendić Lučin Pero Lučin Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection Life cytomegalovirus assembly compartment dynamin early endosomes endosomal recycling compartment Golgi apparatus |
| title | Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection |
| title_full | Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection |
| title_fullStr | Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection |
| title_full_unstemmed | Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection |
| title_short | Dynamin Inhibitors Prevent the Establishment of the Cytomegalovirus Assembly Compartment in the Early Phase of Infection |
| title_sort | dynamin inhibitors prevent the establishment of the cytomegalovirus assembly compartment in the early phase of infection |
| topic | cytomegalovirus assembly compartment dynamin early endosomes endosomal recycling compartment Golgi apparatus |
| url | https://www.mdpi.com/2075-1729/11/9/876 |
| work_keys_str_mv | AT igorstimac dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection AT nataliajugvucko dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection AT gordanablagojeviczagorac dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection AT marinamarcelic dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection AT hanamahmutefendiclucin dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection AT perolucin dynamininhibitorspreventtheestablishmentofthecytomegalovirusassemblycompartmentintheearlyphaseofinfection |