Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport
The intracellular transport of nucleocapsids of the highly pathogenic Marburg, as well as Ebola virus (MARV, EBOV), represents a critical step during the viral life cycle. Intriguingly, a population of these nucleocapsids is distributed over long distances in a directed and polar fashion. Recently,...
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MDPI AG
2020-07-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/7/1728 |
| _version_ | 1797561984543096832 |
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| author | Katharina Grikscheit Olga Dolnik Yuki Takamatsu Ana Raquel Pereira Stephan Becker |
| author_facet | Katharina Grikscheit Olga Dolnik Yuki Takamatsu Ana Raquel Pereira Stephan Becker |
| author_sort | Katharina Grikscheit |
| collection | DOAJ |
| description | The intracellular transport of nucleocapsids of the highly pathogenic Marburg, as well as Ebola virus (MARV, EBOV), represents a critical step during the viral life cycle. Intriguingly, a population of these nucleocapsids is distributed over long distances in a directed and polar fashion. Recently, it has been demonstrated that the intracellular transport of filoviral nucleocapsids depends on actin polymerization. While it was shown that EBOV requires Arp2/3-dependent actin dynamics, the details of how the virus exploits host actin signaling during intracellular transport are largely unknown. Here, we apply a minimalistic transfection system to follow the nucleocapsid-like structures (NCLS) in living cells, which can be used to robustly quantify NCLS transport in live cell imaging experiments. Furthermore, in cells co-expressing LifeAct, a marker for actin dynamics, NCLS transport is accompanied by pulsative actin tails appearing on the rear end of NCLS. These actin tails can also be preserved in fixed cells, and can be visualized via high resolution imaging using STORM in transfected, as well as EBOV infected, cells. The application of inhibitory drugs and siRNA depletion against actin regulators indicated that EBOV NCLS utilize the canonical Arp2/3-Wave1-Rac1 pathway for long-distance transport in cells. These findings highlight the relevance of the regulation of actin polymerization during directed EBOV nucleocapsid transport in human cells. |
| first_indexed | 2024-03-10T18:23:13Z |
| format | Article |
| id | doaj.art-e876e5f793594247b518346ddf829d5e |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T18:23:13Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-e876e5f793594247b518346ddf829d5e2023-11-20T07:16:23ZengMDPI AGCells2073-44092020-07-0197172810.3390/cells9071728Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance TransportKatharina Grikscheit0Olga Dolnik1Yuki Takamatsu2Ana Raquel Pereira3Stephan Becker4Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, GermanyInstitute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, GermanyInstitute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, GermanyOxford Nano Imager, Linacre House, Banbury Rd, Oxford OX2 8TA, UKInstitute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, GermanyThe intracellular transport of nucleocapsids of the highly pathogenic Marburg, as well as Ebola virus (MARV, EBOV), represents a critical step during the viral life cycle. Intriguingly, a population of these nucleocapsids is distributed over long distances in a directed and polar fashion. Recently, it has been demonstrated that the intracellular transport of filoviral nucleocapsids depends on actin polymerization. While it was shown that EBOV requires Arp2/3-dependent actin dynamics, the details of how the virus exploits host actin signaling during intracellular transport are largely unknown. Here, we apply a minimalistic transfection system to follow the nucleocapsid-like structures (NCLS) in living cells, which can be used to robustly quantify NCLS transport in live cell imaging experiments. Furthermore, in cells co-expressing LifeAct, a marker for actin dynamics, NCLS transport is accompanied by pulsative actin tails appearing on the rear end of NCLS. These actin tails can also be preserved in fixed cells, and can be visualized via high resolution imaging using STORM in transfected, as well as EBOV infected, cells. The application of inhibitory drugs and siRNA depletion against actin regulators indicated that EBOV NCLS utilize the canonical Arp2/3-Wave1-Rac1 pathway for long-distance transport in cells. These findings highlight the relevance of the regulation of actin polymerization during directed EBOV nucleocapsid transport in human cells.https://www.mdpi.com/2073-4409/9/7/1728Ebola virusactin cytoskeletonnucleocapsid transportArp2/3 complex |
| spellingShingle | Katharina Grikscheit Olga Dolnik Yuki Takamatsu Ana Raquel Pereira Stephan Becker Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport Cells Ebola virus actin cytoskeleton nucleocapsid transport Arp2/3 complex |
| title | Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport |
| title_full | Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport |
| title_fullStr | Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport |
| title_full_unstemmed | Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport |
| title_short | Ebola Virus Nucleocapsid-Like Structures Utilize Arp2/3 Signaling for Intracellular Long-Distance Transport |
| title_sort | ebola virus nucleocapsid like structures utilize arp2 3 signaling for intracellular long distance transport |
| topic | Ebola virus actin cytoskeleton nucleocapsid transport Arp2/3 complex |
| url | https://www.mdpi.com/2073-4409/9/7/1728 |
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