LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells
Myogenic differentiation is a complex biological process that is regulated by multiple factors, among which long noncoding RNAs (lncRNAs) play an essential role. However, in-depth studies on the regulatory mechanisms of long noncoding RNAs (lncRNAs) in myogenic differentiation are limited. In this s...
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/7/6773 |
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| author | Mengxun Li Quan Liu Su Xie Chong Fu Jiaxuan Li Cheng Tian Xin Li Changchun Li |
| author_facet | Mengxun Li Quan Liu Su Xie Chong Fu Jiaxuan Li Cheng Tian Xin Li Changchun Li |
| author_sort | Mengxun Li |
| collection | DOAJ |
| description | Myogenic differentiation is a complex biological process that is regulated by multiple factors, among which long noncoding RNAs (lncRNAs) play an essential role. However, in-depth studies on the regulatory mechanisms of long noncoding RNAs (lncRNAs) in myogenic differentiation are limited. In this study, we characterized the role of the novel lncRNA <i>TCONS_00323213</i>, which is upregulated during porcine skeletal muscle satellite cell (PSC) differentiation in myogenesis. We found that <i>TCONS_00323213</i> affected the proliferation and differentiation of PSC in vitro. We performed quantitative polymerase chain reaction (qPCR), 5-ethynyl-20-deoxyuridine (EdU), western blotting, immunofluorescence staining, pull-down assays, and cleavage under targets and tagmentation (CUT and Tag) assays to clarify the effects and action mechanisms of <i>TCONS_00323213</i>. LncRNA <i>TCONS_00323213</i> inhibited myoblast proliferation based on analyses of cell survival rates during PSC proliferation. Functional analyses revealed that <i>TCONS_00323213</i> promotes cell differentiation and enhances myogenin (<i>MyoG</i>), myosin heavy chain (<i>MyHC</i>), and myocyte enhancer factor 2 (<i>MEF2C</i>) during myoblast differentiation. As determined by pull-down and RNA immunoprecipitation (RIP) assays, the lncRNA <i>TCONS_00323213</i> interacted with PBX/Knotted Homeobox 2 (PKNOX2). CUT and Tag assays showed that PKNOX2 was significantly enriched on the <i>MyoG</i> promoter after lncRNA <i>TCONS_00323213</i> knockdown. Our findings demonstrate that the interaction between lncRNA <i>TCONS_00323213</i> and PKNOX2 relieves the inhibitory effect of PKNOX2 on the <i>MyoG</i> promoter, increases its expression, and promotes PSC differentiation. This novel role of lncRNA <i>TCONS_00323213</i> sheds light on the molecular mechanisms by which lncRNAs regulate porcine myogenesis. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T05:33:44Z |
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| spelling | doaj.art-51081da7f05041fbb19abe8cc0d408b12023-11-17T16:55:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01247677310.3390/ijms24076773LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite CellsMengxun Li0Quan Liu1Su Xie2Chong Fu3Jiaxuan Li4Cheng Tian5Xin Li6Changchun Li7Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaKey Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, Huazhong Agricultural University, Wuhan 430070, ChinaMyogenic differentiation is a complex biological process that is regulated by multiple factors, among which long noncoding RNAs (lncRNAs) play an essential role. However, in-depth studies on the regulatory mechanisms of long noncoding RNAs (lncRNAs) in myogenic differentiation are limited. In this study, we characterized the role of the novel lncRNA <i>TCONS_00323213</i>, which is upregulated during porcine skeletal muscle satellite cell (PSC) differentiation in myogenesis. We found that <i>TCONS_00323213</i> affected the proliferation and differentiation of PSC in vitro. We performed quantitative polymerase chain reaction (qPCR), 5-ethynyl-20-deoxyuridine (EdU), western blotting, immunofluorescence staining, pull-down assays, and cleavage under targets and tagmentation (CUT and Tag) assays to clarify the effects and action mechanisms of <i>TCONS_00323213</i>. LncRNA <i>TCONS_00323213</i> inhibited myoblast proliferation based on analyses of cell survival rates during PSC proliferation. Functional analyses revealed that <i>TCONS_00323213</i> promotes cell differentiation and enhances myogenin (<i>MyoG</i>), myosin heavy chain (<i>MyHC</i>), and myocyte enhancer factor 2 (<i>MEF2C</i>) during myoblast differentiation. As determined by pull-down and RNA immunoprecipitation (RIP) assays, the lncRNA <i>TCONS_00323213</i> interacted with PBX/Knotted Homeobox 2 (PKNOX2). CUT and Tag assays showed that PKNOX2 was significantly enriched on the <i>MyoG</i> promoter after lncRNA <i>TCONS_00323213</i> knockdown. Our findings demonstrate that the interaction between lncRNA <i>TCONS_00323213</i> and PKNOX2 relieves the inhibitory effect of PKNOX2 on the <i>MyoG</i> promoter, increases its expression, and promotes PSC differentiation. This novel role of lncRNA <i>TCONS_00323213</i> sheds light on the molecular mechanisms by which lncRNAs regulate porcine myogenesis.https://www.mdpi.com/1422-0067/24/7/6773<i>TCONS_00323213</i>PKNOX2porcine skeletal muscle satellite cellsdifferentiationmyogenesis |
| spellingShingle | Mengxun Li Quan Liu Su Xie Chong Fu Jiaxuan Li Cheng Tian Xin Li Changchun Li LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells International Journal of Molecular Sciences <i>TCONS_00323213</i> PKNOX2 porcine skeletal muscle satellite cells differentiation myogenesis |
| title | LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells |
| title_full | LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells |
| title_fullStr | LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells |
| title_full_unstemmed | LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells |
| title_short | LncRNA <i>TCONS_00323213</i> Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate <i>MyoG</i> in Porcine Satellite Cells |
| title_sort | lncrna i tcons 00323213 i promotes myogenic differentiation by interacting with pknox2 to upregulate i myog i in porcine satellite cells |
| topic | <i>TCONS_00323213</i> PKNOX2 porcine skeletal muscle satellite cells differentiation myogenesis |
| url | https://www.mdpi.com/1422-0067/24/7/6773 |
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