N6-methyladenosine is required for efficient RNA synthesis of Ebola virus and other haemorrhagic fever viruses

Emerg Microbes Infect. 2023 Dec;12(2):2223732. doi: 10.1080/22221751.2023.2223732.

Abstract

N6-methyladenosine (m6A) is one of the most abundant modifications of cellular RNA, where it serves various functions. m6A methylation of many viral RNA species has also been described; however, little is known about the m6A epitranscriptome of haemorrhagic fever-causing viruses like Ebola virus (EBOV). Here, we analysed the importance of the methyltransferase METTL3 for the life cycle of this virus. We found that METTL3 interacts with the EBOV nucleoprotein and the transcriptional activator VP30 to support viral RNA synthesis, and that METTL3 is recruited into EBOV inclusions bodies, where viral RNA synthesis occurs. Analysis of the m6A methylation pattern of EBOV mRNAs showed that they are methylated by METTL3. Further studies revealed that METTL3 interaction with the viral nucleoprotein, as well as its importance for RNA synthesis and protein expression, is also observed for other haemorrhagic fever viruses such as Junín virus (JUNV) and Crimean-Congo haemorrhagic fever virus (CCHFV). The negative effects on viral RNA synthesis due to loss of m6A methylation are independent of innate immune sensing, as METTL3 knockout did not affect type I interferon induction in response to viral RNA synthesis or infection. Our results suggest a novel function for m6A that is conserved among diverse haemorrhagic fever-causing viruses (i.e. EBOV, JUNV and CCHFV), making METTL3 a promising target for broadly-acting antivirals.

Keywords: Crimean-Congo haemorrhagic fever virus; Ebola virus; Junín virus; METTL3; arenavirus; filovirus; m6A; orthonairovirus.

MeSH terms

  • Dengue Virus* / genetics
  • Ebolavirus* / genetics
  • Hemorrhagic Fever Virus, Crimean-Congo* / genetics
  • Hemorrhagic Fever, Ebola*
  • Humans
  • Methyltransferases / genetics
  • Nucleoproteins
  • RNA, Viral / genetics
  • RNA, Viral / metabolism

Substances

  • N-methyladenosine
  • RNA, Viral
  • Nucleoproteins
  • METTL3 protein, human
  • Methyltransferases

Grants and funding

Funding was provided by the Deutsche Forschungsgemeinschaft (DFG), grant number 389002253 (L.W.) and grant number 452208680 (B.S.B), as well as by the Friedrich-Loeffler-Institut, intramural funding (S.R., W.F., A.G. and T.H.) and funding as part of the VISION consortium (M.J.P., L.F.). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.