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Evidence for the extracellular delivery of influenza NS1 protein

https://doi.org/10.18527/2500-2236-2021-8-1-27-37

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Аннотация

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.

Об авторах

A. A. Pulkina
Smorodintsev Research Institute of Influenza; Peter the Great St. Petersburg Polytechnic University
Россия

15/17, Prof. Popov str., Saint Petersburg, 197376;
29, Polytechnicheskaya str., Saint Petersburg, 195251



M. V. Sergeeva
Smorodintsev Research Institute of Influenza; Peter the Great St. Petersburg Polytechnic University
Россия

15/17, Prof. Popov str., Saint Petersburg, 197376;
29, Polytechnicheskaya str., Saint Petersburg, 195251



A. Krokhin
Smorodintsev Research Institute of Influenza
Россия

15/17, Prof. Popov str., Saint Petersburg, 197376



M. A. Stukova
Smorodintsev Research Institute of Influenza
Россия

15/17, Prof. Popov str., Saint Petersburg, 197376



A. Egorov
Smorodintsev Research Institute of Influenza
Россия

15/17, Prof. Popov str., Saint Petersburg, 197376



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Для цитирования:


Pulkina A.A., Sergeeva M.V., Krokhin A., Stukova M.A., Egorov A. Evidence for the extracellular delivery of influenza NS1 protein. Microbiology Independent Research Journal (MIR Journal). 2021;8(1):27-37. https://doi.org/10.18527/2500-2236-2021-8-1-27-37

For citation:


Pulkina A.A., Sergeeva M.V., Krokhin A., Stukova M.A., Egorov A. Evidence for the extracellular delivery of influenza NS1 protein. Microbiology Independent Research Journal (MIR Journal). 2021;8(1):27-37. https://doi.org/10.18527/2500-2236-2021-8-1-27-37

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