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. 2021 Jan 7;13(1):75. doi: 10.3390/v13010075

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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ORF4 within gt1 HEV regulated via IRES-like RNA that is upregulated through endoplasmic reticulum (ER) stress. (A) Sequence alignment of the gt1 (GenBank accession no. AF444002) region from 2664 to 2845 bp including IRES sequence compared to homologous sequence of gt3 (GenBank accession no. JQ679013) shows significant sequence homology. (B) IRES-like RNA activity is only seen in gt1 HEV. The ratio of nano luciferase (NL) to firefly luciferase (FFL) is compared in gt1 and gt3 IRES-like RNA element sequences in a dual luciferase reporter plasmid, in the presence of tunicamycin (ER stressor). (C) Translation maps of HEV ORF1 nucleotides encompassing the +1 frameshifted ORF4 reading frame start methionine from gt1 HEV and the comparable region from gt3. Methionine (blue arrow) the translation initiator amino acid of ORF4 within gt1 HEV is not present at the homologous position of the gt3 HEV. Each bar displays the mean of three independent biological experiments with four replicates per sample (** p < 0.01, t-test).

ORF4 within gt1 HEV regulated via IRES-like RNA that is upregulated through endoplasmic reticulum (ER) stress. (A) Sequence alignment of the gt1 (GenBank accession no. AF444002) region from 2664 to 2845 bp including IRES sequence compared to homologous sequence of gt3 (GenBank accession no. JQ679013) shows significant sequence homology. (B) IRES-like RNA activity is only seen in gt1 HEV. The ratio of nano luciferase (NL) to firefly luciferase (FFL) is compared in gt1 and gt3 IRES-like RNA element sequences in a dual luciferase reporter plasmid, in the presence of tunicamycin (ER stressor). (C) Translation maps of HEV ORF1 nucleotides encompassing the +1 frameshifted ORF4 reading frame start methionine from gt1 HEV and the comparable region from gt3. Methionine (blue arrow) the translation initiator amino acid of ORF4 within gt1 HEV is not present at the homologous position of the gt3 HEV. Each bar displays the mean of three independent biological experiments with four replicates per sample (** p < 0.01, t-test).