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m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast

Radhika A. Varier, Theodora Sideri, View ORCID ProfileCharlotte Capitanchik, Zornitsa Manova, Enrica Calvani, View ORCID ProfileAlice Rossi, View ORCID ProfileRaghu R. Edupuganti, Imke Ensinck, Vincent W.C. Chan, View ORCID ProfileHarshil Patel, View ORCID ProfileJoanna Kirkpatrick, View ORCID ProfilePeter Faull, Ambrosius P. Snijders, View ORCID ProfileMichiel Vermeulen, View ORCID ProfileMarkus Ralser, View ORCID ProfileJernej Ule, View ORCID ProfileNicholas M. Luscombe, View ORCID ProfileFolkert J. van Werven
doi: https://doi.org/10.1101/2022.01.20.477035
Radhika A. Varier
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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  • For correspondence: radhikaav@gmail.com folkert.vanwerven@crick.ac.uk
Theodora Sideri
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Charlotte Capitanchik
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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  • ORCID record for Charlotte Capitanchik
Zornitsa Manova
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Enrica Calvani
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Alice Rossi
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Raghu R. Edupuganti
2Department of Molecular Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences (RIMLS), Oncode Institute, Radboud University Nijmegen, 6525 GA Nijmegen, the Netherlands
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  • ORCID record for Raghu R. Edupuganti
Imke Ensinck
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Vincent W.C. Chan
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Harshil Patel
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Joanna Kirkpatrick
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Peter Faull
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
5Biological Mass Spectrometry Facility, 2500 Speedway, The University of Texas at Austin, TX 78712
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Ambrosius P. Snijders
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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Michiel Vermeulen
2Department of Molecular Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences (RIMLS), Oncode Institute, Radboud University Nijmegen, 6525 GA Nijmegen, the Netherlands
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Markus Ralser
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
6Charité Universitätsmedizin Berlin, Department of Biochemistry, Germany
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Jernej Ule
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
3Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology, Queen Square, London WC1N 3BG, UK; National Institute of Chemistry, Hajdrihova 19, 1001 Ljubljana, Slovenia
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Nicholas M. Luscombe
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
4Department of Genetics, Evolution and Environment, UCL Genetics Institute, Gower Street, London WC1E 6BT, UK; Okinawa Institute of Science & Technology Graduate University, 1919-1 Tancha, Onna-son, Kunigami-gun, Okinawa 904-0495, Japan
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Folkert J. van Werven
1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK
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  • ORCID record for Folkert J. van Werven
  • For correspondence: radhikaav@gmail.com folkert.vanwerven@crick.ac.uk
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Abstract

N6-methyladenosine (m6A) RNA modification impacts mRNA fate primarily via reader proteins, which dictate processes in development, stress, and disease. Yet little is known about m6A function in Saccharomyces cerevisiae, which occurs solely during early meiosis. Here we perform a multifaceted analysis of the m6A reader protein Pho92/Mrb1. Cross-linking immunoprecipitation analysis reveals that Pho92 associates with the 3’end of meiotic mRNAs in both an m6A-dependent and independent manner. Within cells, Pho92 transitions from the nucleus to the cytoplasm, and associates with translating ribosomes. In the nucleus Pho92 associates with target loci through its interaction with transcriptional elongator Paf1C. Functionally, we show that Pho92 promotes and links protein synthesis to mRNA decay. As such, the Pho92-mediated m6A-mRNA decay is contingent on active translation and the CCR4-NOT complex. We propose that the m6A reader Pho92 is loaded co-transcriptionally to facilitate correct translation and subsequent decay of m6A modified transcripts, and thereby promotes meiosis.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 21, 2022.
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m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast
Radhika A. Varier, Theodora Sideri, Charlotte Capitanchik, Zornitsa Manova, Enrica Calvani, Alice Rossi, Raghu R. Edupuganti, Imke Ensinck, Vincent W.C. Chan, Harshil Patel, Joanna Kirkpatrick, Peter Faull, Ambrosius P. Snijders, Michiel Vermeulen, Markus Ralser, Jernej Ule, Nicholas M. Luscombe, Folkert J. van Werven
bioRxiv 2022.01.20.477035; doi: https://doi.org/10.1101/2022.01.20.477035
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m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast
Radhika A. Varier, Theodora Sideri, Charlotte Capitanchik, Zornitsa Manova, Enrica Calvani, Alice Rossi, Raghu R. Edupuganti, Imke Ensinck, Vincent W.C. Chan, Harshil Patel, Joanna Kirkpatrick, Peter Faull, Ambrosius P. Snijders, Michiel Vermeulen, Markus Ralser, Jernej Ule, Nicholas M. Luscombe, Folkert J. van Werven
bioRxiv 2022.01.20.477035; doi: https://doi.org/10.1101/2022.01.20.477035

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