Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts
Methyltransferase 3 (METTL3), which has been demonstrated to play a crucial role in a variety of biological processes, is the key enzyme for catalyzing m<sup>6</sup>A modification in RNA. However, the complete protein sequence of METTL3 in quail has not been annotated, and its function i...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-04-01
|
| Series: | Veterinary Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2306-7381/10/4/300 |
| _version_ | 1827743438889549824 |
|---|---|
| author | Jing Liu Wentao Zhang Wei Luo Shuibing Liu Hongxia Jiang Sanfeng Liu Jiguo Xu Biao Chen |
| author_facet | Jing Liu Wentao Zhang Wei Luo Shuibing Liu Hongxia Jiang Sanfeng Liu Jiguo Xu Biao Chen |
| author_sort | Jing Liu |
| collection | DOAJ |
| description | Methyltransferase 3 (METTL3), which has been demonstrated to play a crucial role in a variety of biological processes, is the key enzyme for catalyzing m<sup>6</sup>A modification in RNA. However, the complete protein sequence of METTL3 in quail has not been annotated, and its function in skeletal muscle of quails remains unknown. In the current study, the full-length coding sequence of the quail <i>METTL3</i> was obtained through the 3′ rapid amplification of cDNA ends (3’ RACE) and its homology with that of other species was predicted based on a generated phylogenetic tree. A Cell Counting Kit-8 assay and flow cytometry in a quail myoblast cell line (QM7) demonstrated that METTL3 promotes myoblast proliferation. The overexpression of METTL3 in QM7 cells significantly increased the expression levels of the myoblast differentiation markers myogenin (MYOG), myogenic differentiation 1 (MYOD1), and myocyte enhancer factor 2C (MEF2C), further demonstrating that METTL3 promotes myoblast differentiation. Additionally, transcriptome sequencing following METTL3 overexpression revealed that <i>METTL3</i> controls the expression of various genes involved in RNA splicing and the regulation of gene expression, as well as pathways such as the MAPK signaling pathway. Taken together, our findings demonstrated that <i>METTL3</i> plays a vital function in quail myoblast proliferation and differentiation and that the METTL3-mediated RNA m<sup>6</sup>A modification represents an important epigenetic regulatory mechanism in poultry skeletal muscle development. |
| first_indexed | 2024-03-11T04:27:04Z |
| format | Article |
| id | doaj.art-516d5160b8dd4ff1b3c364df97b786c2 |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-03-11T04:27:04Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-516d5160b8dd4ff1b3c364df97b786c22023-11-17T21:44:01ZengMDPI AGVeterinary Sciences2306-73812023-04-0110430010.3390/vetsci10040300Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail MyoblastsJing Liu0Wentao Zhang1Wei Luo2Shuibing Liu3Hongxia Jiang4Sanfeng Liu5Jiguo Xu6Biao Chen7College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaState Key Laboratory of Livestock and Poultry Breeding, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510000, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaInstitute of Biological Technology, Nanchang Normal University, Nanchang 330032, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaMethyltransferase 3 (METTL3), which has been demonstrated to play a crucial role in a variety of biological processes, is the key enzyme for catalyzing m<sup>6</sup>A modification in RNA. However, the complete protein sequence of METTL3 in quail has not been annotated, and its function in skeletal muscle of quails remains unknown. In the current study, the full-length coding sequence of the quail <i>METTL3</i> was obtained through the 3′ rapid amplification of cDNA ends (3’ RACE) and its homology with that of other species was predicted based on a generated phylogenetic tree. A Cell Counting Kit-8 assay and flow cytometry in a quail myoblast cell line (QM7) demonstrated that METTL3 promotes myoblast proliferation. The overexpression of METTL3 in QM7 cells significantly increased the expression levels of the myoblast differentiation markers myogenin (MYOG), myogenic differentiation 1 (MYOD1), and myocyte enhancer factor 2C (MEF2C), further demonstrating that METTL3 promotes myoblast differentiation. Additionally, transcriptome sequencing following METTL3 overexpression revealed that <i>METTL3</i> controls the expression of various genes involved in RNA splicing and the regulation of gene expression, as well as pathways such as the MAPK signaling pathway. Taken together, our findings demonstrated that <i>METTL3</i> plays a vital function in quail myoblast proliferation and differentiation and that the METTL3-mediated RNA m<sup>6</sup>A modification represents an important epigenetic regulatory mechanism in poultry skeletal muscle development.https://www.mdpi.com/2306-7381/10/4/300quailMETTL3myoblastgene expressionproliferationdifferentiation |
| spellingShingle | Jing Liu Wentao Zhang Wei Luo Shuibing Liu Hongxia Jiang Sanfeng Liu Jiguo Xu Biao Chen Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts Veterinary Sciences quail METTL3 myoblast gene expression proliferation differentiation |
| title | Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts |
| title_full | Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts |
| title_fullStr | Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts |
| title_full_unstemmed | Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts |
| title_short | Cloning of the RNA m<sup>6</sup>A Methyltransferase 3 and Its Impact on the Proliferation and Differentiation of Quail Myoblasts |
| title_sort | cloning of the rna m sup 6 sup a methyltransferase 3 and its impact on the proliferation and differentiation of quail myoblasts |
| topic | quail METTL3 myoblast gene expression proliferation differentiation |
| url | https://www.mdpi.com/2306-7381/10/4/300 |
| work_keys_str_mv | AT jingliu cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT wentaozhang cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT weiluo cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT shuibingliu cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT hongxiajiang cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT sanfengliu cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT jiguoxu cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts AT biaochen cloningofthernamsup6supamethyltransferase3anditsimpactontheproliferationanddifferentiationofquailmyoblasts |