Visualization of RNA virus infection in a marine protist with a universal biomarker
Abstract Half of the marine virosphere is hypothesized to be RNA viruses (kingdom Orthornavirae) that infect abundant micro-eukaryotic hosts (e.g. protists). To test this, quantitative approaches that broadly track infections in situ are needed. Here, we describe a technique—dsRNA-Immunofluorescence...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-31507-w |
| _version_ | 1827966305818378240 |
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| author | Samantha R. Coy Budi Utama James W. Spurlin Julia G. Kim Harshavardhan Deshmukh Peter Lwigale Keizo Nagasaki Adrienne M. S. Correa |
| author_facet | Samantha R. Coy Budi Utama James W. Spurlin Julia G. Kim Harshavardhan Deshmukh Peter Lwigale Keizo Nagasaki Adrienne M. S. Correa |
| author_sort | Samantha R. Coy |
| collection | DOAJ |
| description | Abstract Half of the marine virosphere is hypothesized to be RNA viruses (kingdom Orthornavirae) that infect abundant micro-eukaryotic hosts (e.g. protists). To test this, quantitative approaches that broadly track infections in situ are needed. Here, we describe a technique—dsRNA-Immunofluorescence (dsRIF)—that uses a double-stranded RNA (dsRNA) targeting monoclonal antibody to assess host infection status based on the presence of dsRNA, a replicative intermediate of all Orthornavirae infections. We show that the dinoflagellate Heterocapsa circularisquama produces dsRIF signal ~ 1000 times above background autofluorescence when infected by the + ssRNA virus HcRNAV. dsRNA-positive virocells were detected across > 50% of the 48-h infection cycle and accumulated to represent at least 63% of the population. Photosynthetic and chromosomal integrity remained intact during peak replication, indicating HcRNAV infection does not interrupt these processes. This work validates the use of dsRIF on marine RNA viruses and their hosts, setting the stage for quantitative environmental applications that will accelerate understanding of virus-driven ecosystem impacts. |
| first_indexed | 2024-04-09T17:47:53Z |
| format | Article |
| id | doaj.art-1d5b368db1e4425e890e0fefa62a1788 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-09T17:47:53Z |
| publishDate | 2023-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-1d5b368db1e4425e890e0fefa62a17882023-04-16T11:13:30ZengNature PortfolioScientific Reports2045-23222023-04-0113111210.1038/s41598-023-31507-wVisualization of RNA virus infection in a marine protist with a universal biomarkerSamantha R. Coy0Budi Utama1James W. Spurlin2Julia G. Kim3Harshavardhan Deshmukh4Peter Lwigale5Keizo Nagasaki6Adrienne M. S. Correa7Department of Biosciences, Rice UniversityShared Equipment Authority, Rice UniversityDepartment of Biosciences, Rice UniversityDepartment of Biosciences, Rice UniversityShared Equipment Authority, Rice UniversityDepartment of Biosciences, Rice UniversityFaculty of Science and Technology, Kochi UniversityDepartment of Biosciences, Rice UniversityAbstract Half of the marine virosphere is hypothesized to be RNA viruses (kingdom Orthornavirae) that infect abundant micro-eukaryotic hosts (e.g. protists). To test this, quantitative approaches that broadly track infections in situ are needed. Here, we describe a technique—dsRNA-Immunofluorescence (dsRIF)—that uses a double-stranded RNA (dsRNA) targeting monoclonal antibody to assess host infection status based on the presence of dsRNA, a replicative intermediate of all Orthornavirae infections. We show that the dinoflagellate Heterocapsa circularisquama produces dsRIF signal ~ 1000 times above background autofluorescence when infected by the + ssRNA virus HcRNAV. dsRNA-positive virocells were detected across > 50% of the 48-h infection cycle and accumulated to represent at least 63% of the population. Photosynthetic and chromosomal integrity remained intact during peak replication, indicating HcRNAV infection does not interrupt these processes. This work validates the use of dsRIF on marine RNA viruses and their hosts, setting the stage for quantitative environmental applications that will accelerate understanding of virus-driven ecosystem impacts.https://doi.org/10.1038/s41598-023-31507-w |
| spellingShingle | Samantha R. Coy Budi Utama James W. Spurlin Julia G. Kim Harshavardhan Deshmukh Peter Lwigale Keizo Nagasaki Adrienne M. S. Correa Visualization of RNA virus infection in a marine protist with a universal biomarker Scientific Reports |
| title | Visualization of RNA virus infection in a marine protist with a universal biomarker |
| title_full | Visualization of RNA virus infection in a marine protist with a universal biomarker |
| title_fullStr | Visualization of RNA virus infection in a marine protist with a universal biomarker |
| title_full_unstemmed | Visualization of RNA virus infection in a marine protist with a universal biomarker |
| title_short | Visualization of RNA virus infection in a marine protist with a universal biomarker |
| title_sort | visualization of rna virus infection in a marine protist with a universal biomarker |
| url | https://doi.org/10.1038/s41598-023-31507-w |
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