Loss of the Otx2-Binding Site in the Nanog Promoter Affects the Integrity of Embryonic Stem Cell Subtypes and Specification of Inner Cell Mass-Derived Epiblast.
Mouse embryonic stem cells (ESCs) and the inner cell mass (ICM)-derived epiblast exhibit naive pluripotency. ESC-derived epiblast stem cells (EpiSCs) and the postimplantation epiblast exhibit primed pluripotency. Although core pluripotency factors are well-characterized, additional regulators, including Otx2, recently have been shown to function during the transition from naive to primed pluripotency. Here we uncover a role for Otx2 in the control of the naive pluripotent state. We analyzed Otx2-binding activity in ESCs and EpiSCs and identified Nanog, Oct4, and Sox2 as direct targets. To unravel the Otx2 transcriptional network, we targeted the strongest Otx2-binding site in the Nanog promoter, finding that this site modulates the size of specific ESC-subtype compartments in cultured cells and promotes Nanog expression in vivo, predisposing ICM differentiation to epiblast. Otx2-mediated Nanog regulation thus contributes to the integrity of the ESC state and cell lineage specification in preimplantation development.
Institute of Genetics and Biophysics "Adriano Buzzati-Traverso," CNR, Via P. Castellino 111, 80131 Naples, Italy; IRCCS Neuromed, 86077 Pozzilli (IS), Italy.
Dario
D
Acampora
Institute of Genetics and Biophysics "Adriano Buzzati-Traverso," CNR, Via P. Castellino 111, 80131 Naples, Italy.
Daniela
D
Omodei
Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via Sergio Pansini 5, 80131 Naples, Italy.
Giuseppe
G
Petrosino
Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università degli Studi di Napoli, Via L. De Crecchio, 7, 80138 Naples, Italy; Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei, 34, 80087 Pozzuoli (NA), Italy.
Arcomaria
A
Garofalo
Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università degli Studi di Napoli, Via L. De Crecchio, 7, 80138 Naples, Italy; Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei, 34, 80087 Pozzuoli (NA), Italy.
Marco
M
Savarese
Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università degli Studi di Napoli, Via L. De Crecchio, 7, 80138 Naples, Italy; Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei, 34, 80087 Pozzuoli (NA), Italy.
Vincenzo
V
Nigro
Institute of Genetics and Biophysics "Adriano Buzzati-Traverso," CNR, Via P. Castellino 111, 80131 Naples, Italy.
Luca Giovanni
LG
Di Giovannantonio
Institute of Genetics and Biophysics "Adriano Buzzati-Traverso," CNR, Via P. Castellino 111, 80131 Naples, Italy.
Vincenzo
V
Mercadante
Institute of Genetics and Biophysics "Adriano Buzzati-Traverso," CNR, Via P. Castellino 111, 80131 Naples, Italy; IRCCS Neuromed, 86077 Pozzilli (IS), Italy. Electronic address: antonio.simeone@igb.cnr.it.
Antonio
A
Simeone
D000818:Animals; D001665:Binding Sites; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002451:Cell Compartmentation / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002678:Chimera / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005855:Germ Layers / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D009154:Mutation / Q000235:genetics; D000071317:Nanog Homeobox Protein / Q000235:genetics* / Q000378:metabolism; D051857:Otx Transcription Factors / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000187:drug effects
D053605:Leukemia Inhibitory Factor; D000071317:Nanog Homeobox Protein; D051857:Otx Transcription Factors; C497660:Otx2 protein, mouse
Nanog; Otx2; embryonic stem cells; pluripotency; preimplantation epiblast