Hakai is required for stabilization of core components of the m6A mRNA methylation machinery

Research output: Contribution to journalArticlepeer-review

Authors

  • Praveen Bawankar
  • Tina Lence
  • Chiara Paolantoni
  • Jean-Yves Roignant
  • Migle Kazlauskiene
  • Dominik Jacob
  • Jan B. Heidelberger
  • Florian M Richter
  • Violeta Morin
  • Nastasja Kreim
  • Petra Beli
  • Mark Helm
  • Martin Jinek
  • Jean-Yves Roignant

Colleges, School and Institutes

External organisations

  • Institute of Molecular Biology (IMB), Mainz

Abstract

N6-methyladenosine (m6A) is the most abundant internal modification on mRNA which influences most steps of mRNA metabolism and is involved in several biological functions. The E3 ubiquitin ligase Hakai was previously found in complex with components of the m6A methylation machinery in plants and mammalian cells but its precise function remained to be investigated. Here we show that Hakai is a conserved component of the methyltransferase complex in Drosophila and human cells. In Drosophila, its depletion results in reduced m6A levels and altered m6A-dependent functions including sex determination. We show that its ubiquitination domain is required for dimerization and interaction with other members of the m6A machinery, while its catalytic activity is dispensable. Finally, we demonstrate that the loss of Hakai destabilizes several subunits of the methyltransferase complex, resulting in impaired m6A deposition. Our work adds functional and molecular insights into the mechanism of the m6A mRNA writer complex.

Bibliographic note

Funding Information: We thank the Bloomington, Kyoto and Vienna stock Center for fly lines and the Drosophila Genomics Resource Center at Indiana University for plasmids and cell lines. We are indebted to S. Hayashi for his effort to recover the Hakai1 allele as well as BestGene and the University of Cambridge Department of Genetics Fly Facility for injections. We thank members of the Roignant and Soller lab for helpful discussion. We thank Christian Renz from the Ulrich group at IMB for sharing reagents and advices for the yeast-two-hybrid assay and Catia Igreja and Heike Budde from late Elisa Izaurralde lab for sharing plasmids used in this study. We thank the IMB Genomics core facility for their helpful support and the use of its NextSeq500 (INST 247/870-1 FUGG). Research in the laboratory of J.-Y.R. is supported by University of Lausanne, the Swiss National Science Foundation (310030_197906), the Deutsche Forschungsgemeinschaft RO 4681/9-1, RO 4681/12-1 and RO 4681/13-1. M.S. is funded by the BBSRC (BB/R002932/1) and the Leverhulme Trust. Research in the M.J. laboratory is supported by the Swiss National Competence Center for Research (NCCR) RNA & Disease. M.K. is supported by EMBO (ALTF 1087-2018) and Human Frontier Science Program (LT000248 2019-L) postdoctoral fellowships. M.J. is an International Research Scholar of the Howard Hughes Medical Institute and Vallee Scholar of the Bert L & N Kuggie Vallee Foundation. P.Beli is supported by the Emmy Noether Program (BE 5342/1-1 and BE 5342/1-2). C.P. in the lab of J.-Y.R. is supported by a Boehringer Ingelheim Fonds fellowship. Publisher Copyright: © 2021, The Author(s).

Details

Original languageEnglish
Article number3778
Number of pages15
JournalNature Communications
Volume12
Issue number1
Publication statusPublished - 18 Jun 2021

Keywords

  • Adenosine/analogs & derivatives, Animals, Cell Line, Drosophila Proteins/genetics, Drosophila melanogaster, HeLa Cells, Humans, Methylation, Methyltransferases/genetics, RNA Processing, Post-Transcriptional/genetics, RNA Splicing/genetics, RNA, Messenger/genetics, Ubiquitin-Protein Ligases/genetics