6036ce0507bf5f3cd9f1eaee91a58319 pmc_10177049.pdf cbd604225d97afa775293266a191455553a37399 pmc_10177049.pdf 5f2620ea808aa417ae7e278fefe1e2c32391f336d83733bbddb9be6add9535ac pmc_10177049.pdf Title: The E1a Adenoviral Gene Upregulates the Yamanaka Factors to Induce Partial Cellular Reprogramming Subject: The induction of pluripotency by enforced expression of different sets of genes in somatic cells has been achieved with reprogramming technologies first described by Yamanaka’s group. Methodologies for generating induced pluripotent stem cells are as varied as the combinations of genes used. It has previously been reported that the adenoviral E1a gene can induce the expression of two of the Yamanaka factors (c-Myc and Oct-4) and epigenetic changes. Here, we demonstrate that the E1a-12S over-expression is sufficient to induce pluripotent-like characteristics closely to epiblast stem cells in mouse embryonic fibroblasts through the activation of the pluripotency gene regulatory network. These findings provide not only empirical evidence that the expression of one single factor is sufficient for partial reprogramming but also a potential mechanistic explanation for how viral infection could lead to neoplasia if they are surrounded by the appropriate environment or the right medium, as happens with the tumorogenic niche. Keywords: reprogramming; induced pluripotent stem cells; embryonic stem cells; Yamanaka factors; adenovirus; E1a gene Author: Gracia Mendoza, Rebeca González-Pastor, Juan Miguel Sánchez, Altamira Arce-Cerezo, Miguel Quintanilla, Gema Moreno-Bueno, Anna Pujol, Carolina Belmar-López, Alba de Martino, Efrén Riu, Tristan A. Rodriguez and Pilar Martin-Duque Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sun May 7 11:07:41 2023 CEST ModDate: Sun May 7 11:17:44 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3080341 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YWQWLZ+VnURWPalladioL Type 1 Custom yes yes no 10 0 AIFPQY+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 MBBVOX+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 VPFLAK+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 ZZTQUO+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 PFKOHU+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 46 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 68 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 75 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 85 0 AHOENG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 102 0 AHOENH+PalatinoLinotype CID TrueType Identity-H yes yes yes 105 0 AHOENI+PalatinoLinotype TrueType WinAnsi yes yes no 111 0 AHOENK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 114 0 AHOFNF+PalatinoLinotype TrueType MacRoman yes yes no 117 0 AHOENG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 131 0 AHOENH+PalatinoLinotype CID TrueType Identity-H yes yes yes 134 0 AHOENI+PalatinoLinotype TrueType WinAnsi yes yes no 140 0 AHOENK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 143 0 AHOFNF+PalatinoLinotype TrueType MacRoman yes yes no 146 0 AHOENG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 164 0 AHOENH+PalatinoLinotype CID TrueType Identity-H yes yes yes 167 0 AHOENI+PalatinoLinotype TrueType WinAnsi yes yes no 173 0 AHOENK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 176 0 AHOFNF+PalatinoLinotype TrueType MacRoman yes yes no 179 0 AHOENG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 194 0 AHOENH+PalatinoLinotype CID TrueType Identity-H yes yes yes 197 0 AHOENI+PalatinoLinotype TrueType WinAnsi yes yes no 203 0 AHOENK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 206 0 AHOFNF+PalatinoLinotype TrueType MacRoman yes yes no 209 0 AHOENG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 227 0 AHOENH+PalatinoLinotype CID TrueType Identity-H yes yes yes 230 0 AHOENI+PalatinoLinotype TrueType WinAnsi yes yes no 236 0 AHOENK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 239 0 AHOFNF+PalatinoLinotype TrueType MacRoman yes yes no 242 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2023-12-06 02:35:10 CET RepresentationInformation: pmc_10177049.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-12-02 02:18:44 CET Size: 3080341 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 491 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Cell Culture Destination: subsection.2.1 Item: Title: E1a Retroviral Reprogramming of MEF Oct4-GFP Destination: subsection.2.2 Item: Title: Alkaline Phosphatase Activity and E1a Expression Destination: subsection.2.3 Item: Title: Immunocytochemistry Destination: subsection.2.4 Item: Title: Embryoid Bodies Generation Destination: subsection.2.5 Item: Title: RNA Isolation and qPCR Analysis Destination: subsection.2.6 Item: Title: Gene Expression Profiling Destination: subsection.2.7 Item: Title: GeneChIP Assay Destination: subsection.2.8 Item: Title: In Vivo Studies Destination: subsection.2.9 Children: Item: Title: Teratoma Formation Destination: subsubsection.2.9.1 Item: Title: Chimera Production Destination: subsubsection.2.9.2 Item: Title: Histological Studies Destination: subsection.2.10 Item: Title: Statistical Analyses Destination: subsection.2.11 Item: Title: Results Destination: section.3 Children: Item: Title: Generation and Characterization of iPS-like Cells from MEFs by E1a-12S Overexpression Destination: subsection.3.1 Item: Title: E1a Derived iPS-like Cells Are Similar to mES Cells at the Molecular Level Destination: subsection.3.2 Item: Title: iPS-like Cells Can Be Differentiated into Three Germ Layers In Vitro Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.A. Info: Title: The E1a Adenoviral Gene Upregulates the Yamanaka Factors to Induce Partial Cellular Reprogramming Author: Gracia Mendoza, Rebeca González-Pastor, Juan Miguel Sánchez, Altamira Arce-Cerezo, Miguel Quintanilla, Gema Moreno-Bueno, Anna Pujol, Carolina Belmar-López, Alba de Martino, Efrén Riu, Tristan A. Rodriguez and Pilar Martin-Duque Subject: The induction of pluripotency by enforced expression of different sets of genes in somatic cells has been achieved with reprogramming technologies first described by Yamanaka’s group. Methodologies for generating induced pluripotent stem cells are as varied as the combinations of genes used. It has previously been reported that the adenoviral E1a gene can induce the expression of two of the Yamanaka factors (c-Myc and Oct-4) and epigenetic changes. Here, we demonstrate that the E1a-12S over-expression is sufficient to induce pluripotent-like characteristics closely to epiblast stem cells in mouse embryonic fibroblasts through the activation of the pluripotency gene regulatory network. These findings provide not only empirical evidence that the expression of one single factor is sufficient for partial reprogramming but also a potential mechanistic explanation for how viral infection could lead to neoplasia if they are surrounded by the appropriate environment or the right medium, as happens with the tumorogenic niche. 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