Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML
Abstract Background Protein arginine methylations are crucial post-translational modifications (PTMs) in eukaryotes, playing a significant regulatory role in diverse biological processes. Here, we present our investigation into the detailed arginine methylation pattern of the C-terminal RG-rich regi...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-03-01
|
| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12964-025-02130-1 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850252049894080512 |
|---|---|
| author | Yulun Zhong Rou Zhang Lingzi Lu Huijian Tan Yuyu You Yang Mao Yanqiu Yuan |
| author_facet | Yulun Zhong Rou Zhang Lingzi Lu Huijian Tan Yuyu You Yang Mao Yanqiu Yuan |
| author_sort | Yulun Zhong |
| collection | DOAJ |
| description | Abstract Background Protein arginine methylations are crucial post-translational modifications (PTMs) in eukaryotes, playing a significant regulatory role in diverse biological processes. Here, we present our investigation into the detailed arginine methylation pattern of the C-terminal RG-rich region of METTL14, a key component of the m6A RNA methylation machinery, and its functional implications in biology and disease. Methods Using ETD-based mass spectrometry and in vitro enzyme reactions, we uncover a specific arginine methylation pattern on METTL14. RNA methyltransferase activity assays were used to assess the impact of sDMA on METTL3:METTL14 complex activity. RNA immunoprecipitation was used to evaluate mRNA-m6A reader interactions. MeRIP-seq analysis was used to study the genome-wide effect of METTL14 sDMA on m6A modification in acute myeloid leukemia cells. Results We demonstrate that PRMT5 catalyzes the site-specific symmetric dimethylation at R425 and R445 within the extensively methylated RGG/RG motifs of METTL14. We show a positive regulatory role of symmetric dimethylarginines (sDMA) in the catalytic efficiency of the METTL3:METTL14 complex and m6A-specific gene expression in HEK293T and acute myeloid leukemia cells, potentially through the action of m6A reader protein YTHDF1. In addition, the combined inhibition of METTL3 and PRMT5 further reduces the expression of several m6A substrate genes essential for AML proliferation, suggesting a potential therapeutic strategy for AML treatment. Conclusions The study confirms the coexistence of sDMA and aDMA modifications on METTL14’s RGG/RG motifs, with sDMA at R425 and R445 enhancing METTL3:METTL14’s catalytic efficacy and regulating gene expression through m6A deposition in cancer cells. |
| format | Article |
| id | doaj-art-5fd5c53bcf8a4cc28cd3503c23c9e777 |
| institution | OA Journals |
| issn | 1478-811X |
| language | English |
| publishDate | 2025-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj-art-5fd5c53bcf8a4cc28cd3503c23c9e7772025-08-20T01:57:45ZengBMCCell Communication and Signaling1478-811X2025-03-0123111510.1186/s12964-025-02130-1Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AMLYulun Zhong0Rou Zhang1Lingzi Lu2Huijian Tan3Yuyu You4Yang Mao5Yanqiu Yuan6School of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversitySchool of Pharmaceutical Sciences, Sun Yat-Sen UniversityAbstract Background Protein arginine methylations are crucial post-translational modifications (PTMs) in eukaryotes, playing a significant regulatory role in diverse biological processes. Here, we present our investigation into the detailed arginine methylation pattern of the C-terminal RG-rich region of METTL14, a key component of the m6A RNA methylation machinery, and its functional implications in biology and disease. Methods Using ETD-based mass spectrometry and in vitro enzyme reactions, we uncover a specific arginine methylation pattern on METTL14. RNA methyltransferase activity assays were used to assess the impact of sDMA on METTL3:METTL14 complex activity. RNA immunoprecipitation was used to evaluate mRNA-m6A reader interactions. MeRIP-seq analysis was used to study the genome-wide effect of METTL14 sDMA on m6A modification in acute myeloid leukemia cells. Results We demonstrate that PRMT5 catalyzes the site-specific symmetric dimethylation at R425 and R445 within the extensively methylated RGG/RG motifs of METTL14. We show a positive regulatory role of symmetric dimethylarginines (sDMA) in the catalytic efficiency of the METTL3:METTL14 complex and m6A-specific gene expression in HEK293T and acute myeloid leukemia cells, potentially through the action of m6A reader protein YTHDF1. In addition, the combined inhibition of METTL3 and PRMT5 further reduces the expression of several m6A substrate genes essential for AML proliferation, suggesting a potential therapeutic strategy for AML treatment. Conclusions The study confirms the coexistence of sDMA and aDMA modifications on METTL14’s RGG/RG motifs, with sDMA at R425 and R445 enhancing METTL3:METTL14’s catalytic efficacy and regulating gene expression through m6A deposition in cancer cells.https://doi.org/10.1186/s12964-025-02130-1METTL14Symmetric dimethylarginineN6-methyladenosinePost-translational modificationAcute myeloid leukemia |
| spellingShingle | Yulun Zhong Rou Zhang Lingzi Lu Huijian Tan Yuyu You Yang Mao Yanqiu Yuan Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML Cell Communication and Signaling METTL14 Symmetric dimethylarginine N6-methyladenosine Post-translational modification Acute myeloid leukemia |
| title | Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML |
| title_full | Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML |
| title_fullStr | Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML |
| title_full_unstemmed | Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML |
| title_short | Specific sDMA modifications on the RGG/RG motif of METTL14 regulate its function in AML |
| title_sort | specific sdma modifications on the rgg rg motif of mettl14 regulate its function in aml |
| topic | METTL14 Symmetric dimethylarginine N6-methyladenosine Post-translational modification Acute myeloid leukemia |
| url | https://doi.org/10.1186/s12964-025-02130-1 |
| work_keys_str_mv | AT yulunzhong specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT rouzhang specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT lingzilu specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT huijiantan specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT yuyuyou specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT yangmao specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml AT yanqiuyuan specificsdmamodificationsontherggrgmotifofmettl14regulateitsfunctioninaml |