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. 2022 Sep 22;51(2):806–830. doi: 10.1093/nar/gkac761

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 10. — The heterotypic Zα domain tandem repeat of Hs ZBP1 accumulates in dynamic RNP condensates in CyHV-3 infected cells. (A) Cells infected with the CyHV-3 HsZα1-L-Zα2_ZBP1 recombinant expressing the heterotypic Zα domain tandem repeat of Hs ZBP1 instead of ORF112 were fixed, permeabilized, and treated with the indicated enzymes before immunostaining of dsRNA (α-dsRNA, green signal) and ZBP1 (α-ZBP1, red signal). DNA was stained with Dapi (blue signal). (B) Living cells infected with the CyHV-3 Dendra2-HsZα1-L-Zα2_ZBP1 recombinant expressing the fluorescence Dendra2 protein fused at its C-terminal end to the heterotypic Zα domain tandem repeat of Hs ZBP1 were submitted to green-to-red photoconversion on the indicated ROI (FLAP experiment). Representative pictures collected according to time are presented. (C) CCB cells were coinfected with the indicated CyHV-3 recombinant strains (MOI of 1 PFU/cell for each recombinant). Analysis of coinfected cells revealed that mCherry-ORF112 aa189–280 and Dendra2-HsZα1-L-Zα2_ZBP1 colocalize in RNP condensates. (D) Schematic model linking biochemical properties of Zα domains and the formation of RNP observed in CyHV-3 infected cells.