Fluorescent tagging the NS1 protein in yellow fever virus: Replication-capable viruses which produce the secretory GFP-NS1 fusion protein.

Autor: Syzdykova LR; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: laura.rizabekovna@gmail.com., Binke S; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: stephan.binke@web.de., Keyer VV; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: keer@biocenter.kz., Shevtsov AB; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: shevtsov@biocenter.kz., Zaripov MM; Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 142290, Pushchino, Russian Federation. Electronic address: mizaripov1969@gmail.com., Zhylkibayev AA; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: zhylkibayev@biocenter.kz., Ramanculov EM; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: ramanculov@biocenter.kz., Shustov AV; National Center for Biotechnology, Korgalzhin Hwy 13/5, 010000, Nur-Sultan, Kazakhstan. Electronic address: shustov@biocenter.kz.
Jazyk: angličtina
Zdroj: Virus research [Virus Res] 2021 Mar; Vol. 294, pp. 198291. Date of Electronic Publication: 2020 Dec 31.
DOI: 10.1016/j.virusres.2020.198291
Abstrakt: Yellow fever virus, the prototype in the genus Flavivirus, was used to develop viruses in which the nonstructural protein NS1 is genetically fused to GFP in the context of viruses capable of autonomous replication. The GFP-tagging of NS1 at the amino-terminus appeared possible despite the presence of a small and functionally important domain at the NS1's amino-terminus which can be distorted by such fusing. GFP-tagged NS1 viruses were rescued from DNA-launched molecular clones. The initially produced GFP-tagged NS1 virus was capable of only poor replication. Sequential passages of the virus in cell cultures resulted in the appearance of mutations in GFP, NS4A, NS4B and NS5. The mutations which change amino acid sequences of GFP, NS4A and NS5 have the adaptive effect on the replication of GFP-tagged NS1 viruses. The pattern of GFP-fluorescence indicates that the GFP-NS1 fusion protein is produced into the endoplasmic reticulum. The intracellular GFP-NS1 fusion protein colocalizes with dsRNA. The discovered forms of extracellular GFP-NS1 possibly include tetramers and hexamers.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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