Altered subgenomic RNA abundance provides unique insight into SARS-CoV-2 B.1.1.7/Alpha variant infections

COVID-19 Genomics UK COG-UK Consor, Matthew D. Parker, Hazel Stewart, Ola M. Shehata, Benjamin B. Lindsey, Dhruv R. Shah, Sharon Hsu, Alexander J. Keeley, David G. Partridge, Shay Leary, Alison Cope, Amy State, Katie Johnson, Nasar Ali, Rasha Raghei, Joe Heffer, Nikki Smith, Peijun Zhang, Marta Gallis, Stavroula F. LoukaHailey R. Hornsby, Hatoon Alamri, Max Whiteley, Benjamin H. Foulkes, Stella Christou, Paige Wolverson, Manoj Pohare, Samantha E. Hansford, Luke R. Green, Cariad Evans, Mohammad Raza, Dennis Wang, Andrew E. Firth, James R. Edgar, Silvana Gaudieri, Mark O. Collins, Andrew A. Peden, Thushan de Silva

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Matthew Parker et al. use the ARTIC network tiled amplicon PCR and Oxford Nanopore sequencing of thousands of SARS-CoV-2 samples to detect subgenomic RNA changes in the B.1.1.7 lineage endemic in the UK in late 2020/early 2021. They discovered higher subgenomic RNA in B.1.1.7 compared to previous lineages, and find a noncanonical subgenomic RNA that could encode ORF9b.B.1.1.7 lineage SARS-CoV-2 is more transmissible, leads to greater clinical severity, and results in modest reductions in antibody neutralization. Subgenomic RNA (sgRNA) is produced by discontinuous transcription of the SARS-CoV-2 genome. Applying our tool (periscope) to ARTIC Network Oxford Nanopore Technologies genomic sequencing data from 4400 SARS-CoV-2 positive clinical samples, we show that normalised sgRNA is significantly increased in B.1.1.7 (alpha) infections (n = 879). This increase is seen over the previous dominant lineage in the UK, B.1.177 (n = 943), which is independent of genomic reads, E cycle threshold and days since symptom onset at sampling. A noncanonical sgRNA which could represent ORF9b is found in 98.4% of B.1.1.7 SARS-CoV-2 infections compared with only 13.8% of other lineages, with a 16-fold increase in median sgRNA abundance. We demonstrate that ORF9b protein levels are increased 6-fold in B.1.1.7 compared to a B lineage virus in vitro. We hypothesise that increased ORF9b in B.1.1.7 is a direct consequence of a triple nucleotide mutation in nucleocapsid (28280:GAT > CAT, D3L) creating a transcription regulatory-like sequence complementary to a region 3' of the genomic leader. These findings provide a unique insight into the biology of B.1.1.7 and support monitoring of sgRNA profiles to evaluate emerging potential variants of concern.
Original languageEnglish
Article number666
Number of pages10
JournalCommunications Biology
Volume5
Issue number1
DOIs
Publication statusPublished - 5 Jul 2022

Fingerprint

Dive into the research topics of 'Altered subgenomic RNA abundance provides unique insight into SARS-CoV-2 B.1.1.7/Alpha variant infections'. Together they form a unique fingerprint.

Cite this