Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators
VDACs (voltage-dependent anion-selective channels) are pore-forming proteins of the outer mitochondrial membrane, whose permeability is primarily due to VDACs’ presence. In higher eukaryotes, three isoforms are raised during the evolution: they have the same exon–intron organization, and the protein...
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doaj-f9a05229410546f18eb760eead2085ed2020-11-25T03:56:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-01217388738810.3390/ijms21197388Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulatorsFederica Zinghirino0Xena Giada Pappalardo1Angela Messina2Francesca Guarino3Vito De Pinto4Department of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95123 Catania, ItalyDepartment of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95123 Catania, ItalyDepartment of Biological, Geological and Environmental Sciences, Section of Molecular Biology, University of Catania, Viale A. Doria 6, 95125 Catania, ItalyDepartment of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95123 Catania, ItalyDepartment of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95123 Catania, ItalyVDACs (voltage-dependent anion-selective channels) are pore-forming proteins of the outer mitochondrial membrane, whose permeability is primarily due to VDACs’ presence. In higher eukaryotes, three isoforms are raised during the evolution: they have the same exon–intron organization, and the proteins show the same channel-forming activity. We provide a comprehensive analysis of the three human <i>VDAC</i> genes (<i>VDAC</i>1–3), their expression profiles, promoter activity, and potential transcriptional regulators. VDAC isoforms are broadly but also specifically expressed in various human tissues at different levels, with a predominance of VDAC1 and VDAC2 over VDAC3. However, an RNA-seq cap analysis gene expression (CAGE) approach revealed a higher level of transcription activation of <i>VDAC3</i> gene. We experimentally confirmed this information by reporter assay of <i>VDACs</i> promoter activity. Transcription factor binding sites (TFBSs) distribution in the promoters were investigated. The main regulators common to the three <i>VDAC</i> genes were identified as E2F-myc activator/cell cycle (E2FF), Nuclear respiratory factor 1 (NRF1), Krueppel-like transcription factors (KLFS), E-box binding factors (EBOX) transcription factor family members. All of them are involved in cell cycle and growth, proliferation, differentiation, apoptosis, and metabolism. More transcription factors specific for each <i>VDAC</i> gene isoform were identified, supporting the results in the literature, indicating a general role of VDAC1, as an actor of apoptosis for VDAC2, and the involvement in sex determination and development of VDAC3. For the first time, we propose a comparative analysis of human VDAC promoters to investigate their specific biological functions. Bioinformatics and experimental results confirm the essential role of the VDAC protein family in mitochondrial functionality. Moreover, insights about a specialized function and different regulation mechanisms arise for the three isoform gene.https://www.mdpi.com/1422-0067/21/19/7388VDAC isoformsgene structureexpression profilecore promotertranscription factor binding sitesmitochondrial function |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Federica Zinghirino Xena Giada Pappalardo Angela Messina Francesca Guarino Vito De Pinto |
spellingShingle |
Federica Zinghirino Xena Giada Pappalardo Angela Messina Francesca Guarino Vito De Pinto Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators International Journal of Molecular Sciences VDAC isoforms gene structure expression profile core promoter transcription factor binding sites mitochondrial function |
author_facet |
Federica Zinghirino Xena Giada Pappalardo Angela Messina Francesca Guarino Vito De Pinto |
author_sort |
Federica Zinghirino |
title |
Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators |
title_short |
Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators |
title_full |
Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators |
title_fullStr |
Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators |
title_full_unstemmed |
Is the secret of VDAC Isoforms in their gene regulation? Characterization of human <i>VDAC</i> genes expression profile, promoter activity, and transcriptional regulators |
title_sort |
is the secret of vdac isoforms in their gene regulation? characterization of human <i>vdac</i> genes expression profile, promoter activity, and transcriptional regulators |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2020-10-01 |
description |
VDACs (voltage-dependent anion-selective channels) are pore-forming proteins of the outer mitochondrial membrane, whose permeability is primarily due to VDACs’ presence. In higher eukaryotes, three isoforms are raised during the evolution: they have the same exon–intron organization, and the proteins show the same channel-forming activity. We provide a comprehensive analysis of the three human <i>VDAC</i> genes (<i>VDAC</i>1–3), their expression profiles, promoter activity, and potential transcriptional regulators. VDAC isoforms are broadly but also specifically expressed in various human tissues at different levels, with a predominance of VDAC1 and VDAC2 over VDAC3. However, an RNA-seq cap analysis gene expression (CAGE) approach revealed a higher level of transcription activation of <i>VDAC3</i> gene. We experimentally confirmed this information by reporter assay of <i>VDACs</i> promoter activity. Transcription factor binding sites (TFBSs) distribution in the promoters were investigated. The main regulators common to the three <i>VDAC</i> genes were identified as E2F-myc activator/cell cycle (E2FF), Nuclear respiratory factor 1 (NRF1), Krueppel-like transcription factors (KLFS), E-box binding factors (EBOX) transcription factor family members. All of them are involved in cell cycle and growth, proliferation, differentiation, apoptosis, and metabolism. More transcription factors specific for each <i>VDAC</i> gene isoform were identified, supporting the results in the literature, indicating a general role of VDAC1, as an actor of apoptosis for VDAC2, and the involvement in sex determination and development of VDAC3. For the first time, we propose a comparative analysis of human VDAC promoters to investigate their specific biological functions. Bioinformatics and experimental results confirm the essential role of the VDAC protein family in mitochondrial functionality. Moreover, insights about a specialized function and different regulation mechanisms arise for the three isoform gene. |
topic |
VDAC isoforms gene structure expression profile core promoter transcription factor binding sites mitochondrial function |
url |
https://www.mdpi.com/1422-0067/21/19/7388 |
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