Evidence for the extracellular delivery of influenza NS1 protein
We constructed a reporter influenza A/Puerto Rico/8/1934 virus expressing truncated 124aa N-terminal NS1 protein fused to a luciferase reporter sequence (NanoLuc) without signal peptide. The reproduction activity of the vector correlated well with the luminescent activity in the lysates of infected...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Doctrine
2021-06-01
|
| Series: | Microbiology Independent Research Journal |
| Online Access: | https://www.scienceopen.com/hosted-document?doi=10.18527/2500-2236-2021-8-1-27-37 |
| _version_ | 1797221113406685184 |
|---|---|
| author | Anastasia A. Pulkina Maria V. Sergeeva Artem Krokhin Marina A. Stukova Andrej Egorov |
| author_facet | Anastasia A. Pulkina Maria V. Sergeeva Artem Krokhin Marina A. Stukova Andrej Egorov |
| author_sort | Anastasia A. Pulkina |
| collection | DOAJ |
| description | We constructed a reporter influenza A/Puerto Rico/8/1934 virus expressing truncated 124aa N-terminal NS1 protein fused to a luciferase reporter sequence (NanoLuc) without signal peptide. The reproduction activity of the vector correlated well with the luminescent activity in the lysates of infected cell cultures or mouse respiratory organ suspensions. Surprisingly, we found that luciferase enzymatic activity was present not only in the intracellular compartments but also in cell culture supernatants as well as in the sera or bronchiolar lavages of infected mice. This fact allowed us to formulate a working hypothesis about the extracellular delivery mechanism of the NS1 protein. To test this idea, we conducted co-transfection experiments in Vero cells with different combinations of plasmids encoding influenza genomic segments and chimeric NS1-NanoLuc encoding plasmid. We found that the emergence of the luciferase reporter in the extracellular compartment was promoted by the formation of the ribonucleoprotein complex (RNP) from the co-transfection of plasmids expressing PB1, PB2, PA, and NP proteins. Therefore, influenza NS1 protein may be delivered to the extracellular compartment together with the nascent RNP complexes during the maturation of virus particles. |
| first_indexed | 2024-03-11T07:17:07Z |
| format | Article |
| id | doaj.art-ee55ea00b3294c9e92f2df65a0f243db |
| institution | Directory Open Access Journal |
| issn | 2500-2236 |
| language | English |
| last_indexed | 2024-04-24T13:00:16Z |
| publishDate | 2021-06-01 |
| publisher | Doctrine |
| record_format | Article |
| series | Microbiology Independent Research Journal |
| spelling | doaj.art-ee55ea00b3294c9e92f2df65a0f243db2024-04-05T16:00:22ZengDoctrineMicrobiology Independent Research Journal2500-22362021-06-0181273710.18527/2500-2236-2021-8-1-27-37Evidence for the extracellular delivery of influenza NS1 proteinAnastasia A. PulkinaMaria V. SergeevaArtem KrokhinMarina A. StukovaAndrej EgorovWe constructed a reporter influenza A/Puerto Rico/8/1934 virus expressing truncated 124aa N-terminal NS1 protein fused to a luciferase reporter sequence (NanoLuc) without signal peptide. The reproduction activity of the vector correlated well with the luminescent activity in the lysates of infected cell cultures or mouse respiratory organ suspensions. Surprisingly, we found that luciferase enzymatic activity was present not only in the intracellular compartments but also in cell culture supernatants as well as in the sera or bronchiolar lavages of infected mice. This fact allowed us to formulate a working hypothesis about the extracellular delivery mechanism of the NS1 protein. To test this idea, we conducted co-transfection experiments in Vero cells with different combinations of plasmids encoding influenza genomic segments and chimeric NS1-NanoLuc encoding plasmid. We found that the emergence of the luciferase reporter in the extracellular compartment was promoted by the formation of the ribonucleoprotein complex (RNP) from the co-transfection of plasmids expressing PB1, PB2, PA, and NP proteins. Therefore, influenza NS1 protein may be delivered to the extracellular compartment together with the nascent RNP complexes during the maturation of virus particles.https://www.scienceopen.com/hosted-document?doi=10.18527/2500-2236-2021-8-1-27-37 |
| spellingShingle | Anastasia A. Pulkina Maria V. Sergeeva Artem Krokhin Marina A. Stukova Andrej Egorov Evidence for the extracellular delivery of influenza NS1 protein Microbiology Independent Research Journal |
| title | Evidence for the extracellular delivery of influenza NS1 protein |
| title_full | Evidence for the extracellular delivery of influenza NS1 protein |
| title_fullStr | Evidence for the extracellular delivery of influenza NS1 protein |
| title_full_unstemmed | Evidence for the extracellular delivery of influenza NS1 protein |
| title_short | Evidence for the extracellular delivery of influenza NS1 protein |
| title_sort | evidence for the extracellular delivery of influenza ns1 protein |
| url | https://www.scienceopen.com/hosted-document?doi=10.18527/2500-2236-2021-8-1-27-37 |
| work_keys_str_mv | AT anastasiaapulkina evidencefortheextracellulardeliveryofinfluenzans1protein AT mariavsergeeva evidencefortheextracellulardeliveryofinfluenzans1protein AT artemkrokhin evidencefortheextracellulardeliveryofinfluenzans1protein AT marinaastukova evidencefortheextracellulardeliveryofinfluenzans1protein AT andrejegorov evidencefortheextracellulardeliveryofinfluenzans1protein |