<i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage
Promoter-associated noncoding RNAs (pancRNAs) represent a class of noncoding transcripts driven from the promoter region of protein-coding or non-coding genes that operate as cis-acting elements to regulate the expression of the host gene. PancRNAs act by altering the chromatin structure and recruit...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/6/1537 |
| _version_ | 1797446917292032000 |
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| author | Ramona Palombo Maria Paola Paronetto |
| author_facet | Ramona Palombo Maria Paola Paronetto |
| author_sort | Ramona Palombo |
| collection | DOAJ |
| description | Promoter-associated noncoding RNAs (pancRNAs) represent a class of noncoding transcripts driven from the promoter region of protein-coding or non-coding genes that operate as cis-acting elements to regulate the expression of the host gene. PancRNAs act by altering the chromatin structure and recruiting transcription regulators. <i>PncCCND1_B</i> is driven by the promoter region of <i>CCND1</i> and regulates <i>CCND1</i> expression in Ewing sarcoma through recruitment of a multi-molecular complex composed of the RNA binding protein Sam68 and the DNA/RNA helicase DHX9. In this study, we investigated the regulation of <i>CCND1</i> expression in Ewing sarcoma cells upon exposure to chemotherapeutic drugs. Pan-inhibitor screening indicated that etoposide, a drug used for Ewing sarcoma treatment, promotes transcription of <i>pncCCND1_B</i> and repression of <i>CCND1</i> expression. RNA immunoprecipitation experiments showed increased binding of Sam68 to the <i>pncCCND1_B</i> after treatment, despite the significant reduction in DHX9 protein. This effect was associated with the formation of DNA:RNA duplexes at the <i>CCND1</i> promoter. Furthermore, Sam68 interacted with HDAC1 in etoposide treated cells, thus contributing to chromatin remodeling and epigenetic changes. Interestingly, inhibition of the ATM signaling pathway by KU 55,933 treatment was sufficient to inhibit etoposide-induced Sam68-HDAC1 interaction without rescuing DHX9 expression. In these conditions, the DNA:RNA hybrids persist, thus contributing to the local chromatin inactivation at the <i>CCND1</i> promoter region. Altogether, our results show an active role of Sam68 in DNA damage signaling and chromatin remodeling on the <i>CCND1</i> gene by fine-tuning transitions of epigenetic complexes on the <i>CCND1</i> promoter. |
| first_indexed | 2024-03-09T13:47:29Z |
| format | Article |
| id | doaj.art-c1dec3da18a446fca214b37fe07f1491 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T13:47:29Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-c1dec3da18a446fca214b37fe07f14912023-11-30T20:56:48ZengMDPI AGCancers2072-66942022-03-01146153710.3390/cancers14061537<i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA DamageRamona Palombo0Maria Paola Paronetto1Laboratory of Molecular and Cellular Neurobiology, IRCCS Fondazione Santa Lucia, 00143 Rome, ItalyLaboratory of Molecular and Cellular Neurobiology, IRCCS Fondazione Santa Lucia, 00143 Rome, ItalyPromoter-associated noncoding RNAs (pancRNAs) represent a class of noncoding transcripts driven from the promoter region of protein-coding or non-coding genes that operate as cis-acting elements to regulate the expression of the host gene. PancRNAs act by altering the chromatin structure and recruiting transcription regulators. <i>PncCCND1_B</i> is driven by the promoter region of <i>CCND1</i> and regulates <i>CCND1</i> expression in Ewing sarcoma through recruitment of a multi-molecular complex composed of the RNA binding protein Sam68 and the DNA/RNA helicase DHX9. In this study, we investigated the regulation of <i>CCND1</i> expression in Ewing sarcoma cells upon exposure to chemotherapeutic drugs. Pan-inhibitor screening indicated that etoposide, a drug used for Ewing sarcoma treatment, promotes transcription of <i>pncCCND1_B</i> and repression of <i>CCND1</i> expression. RNA immunoprecipitation experiments showed increased binding of Sam68 to the <i>pncCCND1_B</i> after treatment, despite the significant reduction in DHX9 protein. This effect was associated with the formation of DNA:RNA duplexes at the <i>CCND1</i> promoter. Furthermore, Sam68 interacted with HDAC1 in etoposide treated cells, thus contributing to chromatin remodeling and epigenetic changes. Interestingly, inhibition of the ATM signaling pathway by KU 55,933 treatment was sufficient to inhibit etoposide-induced Sam68-HDAC1 interaction without rescuing DHX9 expression. In these conditions, the DNA:RNA hybrids persist, thus contributing to the local chromatin inactivation at the <i>CCND1</i> promoter region. Altogether, our results show an active role of Sam68 in DNA damage signaling and chromatin remodeling on the <i>CCND1</i> gene by fine-tuning transitions of epigenetic complexes on the <i>CCND1</i> promoter.https://www.mdpi.com/2072-6694/14/6/1537Sam68noncoding RNAEwing sarcoma<i>CCND1</i>DNA damage |
| spellingShingle | Ramona Palombo Maria Paola Paronetto <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage Cancers Sam68 noncoding RNA Ewing sarcoma <i>CCND1</i> DNA damage |
| title | <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage |
| title_full | <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage |
| title_fullStr | <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage |
| title_full_unstemmed | <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage |
| title_short | <i>pncCCND1_B</i> Engages an Inhibitory Protein Network to Downregulate <i>CCND1</i> Expression upon DNA Damage |
| title_sort | i pncccnd1 b i engages an inhibitory protein network to downregulate i ccnd1 i expression upon dna damage |
| topic | Sam68 noncoding RNA Ewing sarcoma <i>CCND1</i> DNA damage |
| url | https://www.mdpi.com/2072-6694/14/6/1537 |
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