Details
| Original language | English |
|---|---|
| Pages (from-to) | 1511-1522 |
| Number of pages | 12 |
| Journal | The plant cell |
| Volume | 30 |
| Issue number | 7 |
| Early online date | 8 Jun 2018 |
| Publication status | Published - Jul 2018 |
Abstract
N 6-methylated adenine (m 6A) is the most frequent posttranscriptional modification in eukaryotic mRNA. Turnover of RNA generates N 6-methylated AMP (N 6-mAMP), which has an unclear metabolic fate. We show that Arabidopsis thaliana and human cells require an N 6-mAMP deaminase (ADAL, renamed MAPDA) to catabolize N 6-mAMP to inosine monophosphate in vivo by hydrolytically removing the aminomethyl group. A phylogenetic, structural, and biochemical analysis revealed that many fungi partially or fully lack MAPDA, which coincides with a minor role of N 6A-RNA methylation in these organisms. MAPDA likely protects RNA from m 6A misincorporation. This is required because eukaryotic RNA polymerase can use N 6-mATP as a substrate. Upon abrogation of MAPDA, root growth is slightly reduced, and the N 6-methyladenosine, N 6-mAMP, and N 6-mATP concentrations are increased in Arabidopsis. Although this will potentially lead to m 6A misincorporation into RNA, we show that the frequency is too low to be reliably detected in vivo. Since N 6-mAMP was severalfold more abundant than N 6-mATP in MAPDA mutants, we speculate that additional molecular filters suppress the generation of N 6-mATP. Enzyme kinetic data indicate that adenylate kinases represent such filters being highly selective for AMP versus N 6-mAMP phosphorylation. We conclude that a multilayer molecular protection system is in place preventing N 6-mAMP accumulation and salvage.
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Plant Science
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: The plant cell, Vol. 30, No. 7, 07.2018, p. 1511-1522.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - m6A RNA Degradation Products Are Catabolized by an Evolutionarily Conserved N6-Methyl-AMP Deaminase in Plant and Mammalian Cells
AU - Chen, Mingjia
AU - Urs, Mounashree J.
AU - Sánchez-González, Ismael
AU - Olayioye, Monilola A.
AU - Herde, Marco
AU - Witte, Claus-Peter
N1 - Funding information: We thank André Specht, Hildegard Thölke, and Philipp Rüter for technical support, Xiaoye Liu for help with the statistical analyses, Anting Zhu for assistance with mass spectrometry, and Markus Niehaus for his help with root phenotype analysis. We also thank Rico M. Hartmann, Marian Uhe, and Helge Küster for support with the bioanalyzer. This work was financially supported by the Deutsche Forschungsgemeinschaft (Grants WI3411/2-1, WI3411/4-1, and HE 5949/3-1) and the Leibniz University Hannover (Wege in die Forschung II to M.C.).
PY - 2018/7
Y1 - 2018/7
N2 - N 6-methylated adenine (m 6A) is the most frequent posttranscriptional modification in eukaryotic mRNA. Turnover of RNA generates N 6-methylated AMP (N 6-mAMP), which has an unclear metabolic fate. We show that Arabidopsis thaliana and human cells require an N 6-mAMP deaminase (ADAL, renamed MAPDA) to catabolize N 6-mAMP to inosine monophosphate in vivo by hydrolytically removing the aminomethyl group. A phylogenetic, structural, and biochemical analysis revealed that many fungi partially or fully lack MAPDA, which coincides with a minor role of N 6A-RNA methylation in these organisms. MAPDA likely protects RNA from m 6A misincorporation. This is required because eukaryotic RNA polymerase can use N 6-mATP as a substrate. Upon abrogation of MAPDA, root growth is slightly reduced, and the N 6-methyladenosine, N 6-mAMP, and N 6-mATP concentrations are increased in Arabidopsis. Although this will potentially lead to m 6A misincorporation into RNA, we show that the frequency is too low to be reliably detected in vivo. Since N 6-mAMP was severalfold more abundant than N 6-mATP in MAPDA mutants, we speculate that additional molecular filters suppress the generation of N 6-mATP. Enzyme kinetic data indicate that adenylate kinases represent such filters being highly selective for AMP versus N 6-mAMP phosphorylation. We conclude that a multilayer molecular protection system is in place preventing N 6-mAMP accumulation and salvage.
AB - N 6-methylated adenine (m 6A) is the most frequent posttranscriptional modification in eukaryotic mRNA. Turnover of RNA generates N 6-methylated AMP (N 6-mAMP), which has an unclear metabolic fate. We show that Arabidopsis thaliana and human cells require an N 6-mAMP deaminase (ADAL, renamed MAPDA) to catabolize N 6-mAMP to inosine monophosphate in vivo by hydrolytically removing the aminomethyl group. A phylogenetic, structural, and biochemical analysis revealed that many fungi partially or fully lack MAPDA, which coincides with a minor role of N 6A-RNA methylation in these organisms. MAPDA likely protects RNA from m 6A misincorporation. This is required because eukaryotic RNA polymerase can use N 6-mATP as a substrate. Upon abrogation of MAPDA, root growth is slightly reduced, and the N 6-methyladenosine, N 6-mAMP, and N 6-mATP concentrations are increased in Arabidopsis. Although this will potentially lead to m 6A misincorporation into RNA, we show that the frequency is too low to be reliably detected in vivo. Since N 6-mAMP was severalfold more abundant than N 6-mATP in MAPDA mutants, we speculate that additional molecular filters suppress the generation of N 6-mATP. Enzyme kinetic data indicate that adenylate kinases represent such filters being highly selective for AMP versus N 6-mAMP phosphorylation. We conclude that a multilayer molecular protection system is in place preventing N 6-mAMP accumulation and salvage.
UR - http://www.scopus.com/inward/record.url?scp=85055554583&partnerID=8YFLogxK
U2 - 10.1105/tpc.18.00236
DO - 10.1105/tpc.18.00236
M3 - Article
VL - 30
SP - 1511
EP - 1522
JO - The plant cell
JF - The plant cell
SN - 1040-4651
IS - 7
ER -