Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Hybrid Sequencing Reveals Novel Features in the Transcriptomic Organization of Equid Alphaherpesvirus 1

Version 1 : Received: 14 June 2022 / Approved: 15 June 2022 / Online: 15 June 2022 (09:30:44 CEST)

How to cite: Tombácz, D.; Torma, G.; Gulyás, G.; Fülöp, Á.; Dörmő, Á.; Prazsák, I.; Csabai, Z.; Mizik, M.; Hornyák, Á.; Zoltán, Z.; Kakuk, B.; Boldogkői, Z. Hybrid Sequencing Reveals Novel Features in the Transcriptomic Organization of Equid Alphaherpesvirus 1. Preprints 2022, 2022060219. https://doi.org/10.20944/preprints202206.0219.v1. Tombácz, D.; Torma, G.; Gulyás, G.; Fülöp, Á.; Dörmő, Á.; Prazsák, I.; Csabai, Z.; Mizik, M.; Hornyák, Á.; Zoltán, Z.; Kakuk, B.; Boldogkői, Z. Hybrid Sequencing Reveals Novel Features in the Transcriptomic Organization of Equid Alphaherpesvirus 1. Preprints 2022, 2022060219. https://doi.org/10.20944/preprints202206.0219.v1.

Abstract

In this study, a structural profiling of equid alphaherpesvirus 1 (EHV-1) transcriptome was carried out using next-generation (Illumina) and third-generation (Oxford Nanopore Technologies) sequencing platforms. We annotated the canonical mRNA molecules and their isoforms, including transcript start and end site isoforms, and splice variants. Additionally, a number of putative 5-truncated mRNAs containing shorter in-frame ORFs were detected. We also demonstrated that EHV-1 produces a high number of non-coding transcripts, including antisense and intergenic RNAs. One of the most remarkable features of the EHV-1 is the generation of abundant fusion transcripts some of which encoding chimeric polypeptides. We observed a higher number of splicing and transcriptional overlaps than in related viruses. Additionally, we found that many upstream genes of tandem gene clusters have their own transcript end sites (TESs) besides the co-terminal TESs, which is rare in other alphaherpesviruses. We show here that the replication origins (OriS and OriL) of the virus are co-localized with promoter sequences and overlap with specific RNA molecules. Furthermore, we discovered a novel non-coding RNA (designated as NOIR) that overlaps the 5′-ends of the longer transcript variants encoded by the two main transactivator genes ORF64 and 65 bracketing the OriL. These all suggest the existence of a central regulatory system which controls the genome-wide transcription and the replication through a mechanism based on the interference between the machineries carrying out the synthesis of DNA and RNA.

Keywords

Equid alphaherpesvirus 1; EHV-1; transcriptome; replication origin; long-read sequencing; nanopore sequencing; direct RNA sequencing; Illumina sequencing

Subject

BIOLOGY, Other

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