Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway
Recent evidence suggested that N6-methyladenosine (m<sup>6</sup>A) methylation can determine m<sup>6</sup>A-modified mRNA fate and play an important role in skeletal muscle development. It was well known that transforming growth factor beta 1 (TGFβ1) is involved in a variety...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-03-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/12/7/1005 |
| _version_ | 1797608185826115584 |
|---|---|
| author | Kaiping Deng Zhipeng Liu Xiaodan Li Zhen Zhang Yixuan Fan Qunhao Huang Yanli Zhang Feng Wang |
| author_facet | Kaiping Deng Zhipeng Liu Xiaodan Li Zhen Zhang Yixuan Fan Qunhao Huang Yanli Zhang Feng Wang |
| author_sort | Kaiping Deng |
| collection | DOAJ |
| description | Recent evidence suggested that N6-methyladenosine (m<sup>6</sup>A) methylation can determine m<sup>6</sup>A-modified mRNA fate and play an important role in skeletal muscle development. It was well known that transforming growth factor beta 1 (TGFβ1) is involved in a variety of cellular processes, such as proliferation, differentiation, and apoptosis. However, little is known about the m<sup>6</sup>A-mediated TGFβ1 regulation in myogenesis. Here, we observed an increase in endogenous TGFβ1 expression and activity during myotube differentiation. However, the knockdown of TGFβ1 inhibits the proliferation and induces cell apoptosis of myoblast. Moreover, we found that m<sup>6</sup>A in 5′-untranslated regions (5′UTR) of TGFβ1 promote its decay and inhibit its expression, leading to the blockage of the TGFβ1/SMAD2 signaling pathway. Furthermore, the targeted specific demethylation of TGFβ1 m<sup>6</sup>A using dCas13b-FTO significantly increased the TGFβ1-mediated activity of the SMAD2 signaling pathway, promoting myoblast proliferation. These findings suggest that TGFβ1 is an essential regulator of myoblast growth that is negatively regulated by m<sup>6</sup>A. Overall, these results highlight the critical role of m<sup>6</sup>A-mediated post-transcriptional regulation in myogenesis. |
| first_indexed | 2024-03-11T05:40:48Z |
| format | Article |
| id | doaj.art-703ddba947564d0da2e646fd4167a647 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T05:40:48Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-703ddba947564d0da2e646fd4167a6472023-11-17T16:27:56ZengMDPI AGCells2073-44092023-03-01127100510.3390/cells12071005Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling PathwayKaiping Deng0Zhipeng Liu1Xiaodan Li2Zhen Zhang3Yixuan Fan4Qunhao Huang5Yanli Zhang6Feng Wang7Institute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaAnimal Husbandry and Veterinary Station of Haimen District, Nantong 226100, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaInstitute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing 210095, ChinaRecent evidence suggested that N6-methyladenosine (m<sup>6</sup>A) methylation can determine m<sup>6</sup>A-modified mRNA fate and play an important role in skeletal muscle development. It was well known that transforming growth factor beta 1 (TGFβ1) is involved in a variety of cellular processes, such as proliferation, differentiation, and apoptosis. However, little is known about the m<sup>6</sup>A-mediated TGFβ1 regulation in myogenesis. Here, we observed an increase in endogenous TGFβ1 expression and activity during myotube differentiation. However, the knockdown of TGFβ1 inhibits the proliferation and induces cell apoptosis of myoblast. Moreover, we found that m<sup>6</sup>A in 5′-untranslated regions (5′UTR) of TGFβ1 promote its decay and inhibit its expression, leading to the blockage of the TGFβ1/SMAD2 signaling pathway. Furthermore, the targeted specific demethylation of TGFβ1 m<sup>6</sup>A using dCas13b-FTO significantly increased the TGFβ1-mediated activity of the SMAD2 signaling pathway, promoting myoblast proliferation. These findings suggest that TGFβ1 is an essential regulator of myoblast growth that is negatively regulated by m<sup>6</sup>A. Overall, these results highlight the critical role of m<sup>6</sup>A-mediated post-transcriptional regulation in myogenesis.https://www.mdpi.com/2073-4409/12/7/1005m<sup>6</sup>AdCas13bTGFβ1cell proliferationmyoblast |
| spellingShingle | Kaiping Deng Zhipeng Liu Xiaodan Li Zhen Zhang Yixuan Fan Qunhao Huang Yanli Zhang Feng Wang Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway Cells m<sup>6</sup>A dCas13b TGFβ1 cell proliferation myoblast |
| title | Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway |
| title_full | Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway |
| title_fullStr | Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway |
| title_full_unstemmed | Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway |
| title_short | Targeted Demethylation of the TGFβ1 mRNA Promotes Myoblast Proliferation via Activating the SMAD2 Signaling Pathway |
| title_sort | targeted demethylation of the tgfβ1 mrna promotes myoblast proliferation via activating the smad2 signaling pathway |
| topic | m<sup>6</sup>A dCas13b TGFβ1 cell proliferation myoblast |
| url | https://www.mdpi.com/2073-4409/12/7/1005 |
| work_keys_str_mv | AT kaipingdeng targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT zhipengliu targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT xiaodanli targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT zhenzhang targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT yixuanfan targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT qunhaohuang targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT yanlizhang targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway AT fengwang targeteddemethylationofthetgfb1mrnapromotesmyoblastproliferationviaactivatingthesmad2signalingpathway |