Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 117, Issue 35, Pages 21785-21795Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.2003733117
Keywords
miRNA biogenesis; MTA; m(6)A methylation
Categories
Funding
- Polish National Science Centre [UMO-2019/32/T/NZ1/00122, UMO-2017/27/N/NZ1/00202, UMO-2016/23/B/NZ9/00862, UMO-2016/23/D/NZ1/00152, UMO-2013/10/A/NZ1/00557]
- Initiative of ExcellenceResearch University at Adam Mickiewicz University, Poznan, Poland [05/IDUB/2019/94]
- Poznan RNA Research Centre
- Biotechnology and Biological Sciences Research Council [BB/M008606/1]
- National Science Foundation [MCB-1623887, IOS-1444490]
- BBSRC [BB/M008606/1] Funding Source: UKRI
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In Arabidopsis thaliana, the METTL3 homolog, mRNA adenosine methylase (MTA) introduces N-6-methyladenosine (m(6)A) into various coding and noncoding RNAs of the plant transcriptome. Here, we show that an MTA-deficient mutant (mta) has decreased levels of microRNAs (miRNAs) but accumulates primary miRNA transcripts (pri-miRNAs). Moreover, pri-miRNAs are methylated by MTA, and RNA structure probing analysis reveals a decrease in secondary structure within stem-loop regions of these transcripts in mta mutant plants. We demonstrate interaction between MTA and both RNA Polymerase II and TOUGH (TGH), a plant protein needed for early steps of miRNA biogenesis. Both MTA and TGH are necessary for efficient colocalization of the Microprocessor components Dicer-like 1 (DCL1) and Hyponastic Leaves 1 (HYL1) with RNA Polymerase II. We propose that secondary structure of miRNA precursors induced by their MTA-dependent m(6)A methylation status, together with direct interactions between MTA and TGH, influence the recruitment of Microprocessor to plant pri-miRNAs. Therefore, the lack of MTA in mta mutant plants disturbs pri-miRNA processing and leads to the decrease in miRNA accumulation. Furthermore, our findings reveal that reduced miR393b levels likely contributes to the impaired auxin response phenotypes of mta mutant plants.
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