Modular organization of a hypocretin gene minimal promoter

Adriana Sánchez-García, Griselda A. Cabral-Pacheco, Viviana C. Zomosa-Signoret, Rocío Ortiz-López, Alberto Camacho, Paulo M. Tabera-Tarello, José A. Garnica-López, Román Vidaltamayo

Resultado de la investigación

1 Cita (Scopus)

Resumen

Orexins or hypocretins are neurotransmitters produced by a small population of neurons in the lateral hypothalamus. This family of peptides modulates sleep-wake cycle, arousal and feeding behaviors; however, the mechanisms regulating their expression remain to be fully elucidated. There is an interest in defining the key molecular elements in orexin regulation, as these may serve to identify targets for generating novel therapies for sleep disorders, obesity and addiction. Our previous studies showed that the expression of orexin was decreased in mice carrying null-mutations of the transcription factor early B-cell factor 2 (ebf2) and that the promoter region of the prepro-orexin (Hcrt) gene contained two putative ebf-binding sites, termed olf-1 sites. In the present study, a minimal promoter region of the murine Hcrt gene was identified, which was able to drive the expression of a luciferase reporter gene in the human 293 cell line. Deletion of the olf1-site proximal to the transcription start site of the Hcrt gene increased reporter gene expression, whereas deletion of the distal olf1-like site decreased its expression. The lentiviral transduction of murine transcription factor ebf2 cDNA into 293 cells increased the gene expression driven by this minimal Hcrt-gene promoter and an electrophoretic mobility shift assays demonstrated that the distal olf1-like sequence was a binding site for ebf2.

Idioma originalEnglish
Páginas (desde-hasta)2263-2270
Número de páginas8
PublicaciónMolecular Medicine Reports
Volumen17
N.º2
DOI
EstadoPublished - 1 feb 2018

Huella dactilar

Genes
B-Lymphocytes
Reporter Genes
Genetic Promoter Regions
Transcription Factors
Gene expression
Binding Sites
Lateral Hypothalamic Area
Gene Expression
Cells
Transcription Initiation Site
Electrophoretic Mobility Shift Assay
Feeding Behavior
Arousal
Luciferases
Electrophoretic mobility
Neurotransmitter Agents
Sleep
Complementary DNA
Obesity

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Oncology
  • Cancer Research

Citar esto

Sánchez-García, A., Cabral-Pacheco, G. A., Zomosa-Signoret, V. C., Ortiz-López, R., Camacho, A., Tabera-Tarello, P. M., ... Vidaltamayo, R. (2018). Modular organization of a hypocretin gene minimal promoter. Molecular Medicine Reports, 17(2), 2263-2270. https://doi.org/10.3892/mmr.2017.8142
Sánchez-García, Adriana ; Cabral-Pacheco, Griselda A. ; Zomosa-Signoret, Viviana C. ; Ortiz-López, Rocío ; Camacho, Alberto ; Tabera-Tarello, Paulo M. ; Garnica-López, José A. ; Vidaltamayo, Román. / Modular organization of a hypocretin gene minimal promoter. En: Molecular Medicine Reports. 2018 ; Vol. 17, N.º 2. pp. 2263-2270.
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abstract = "Orexins or hypocretins are neurotransmitters produced by a small population of neurons in the lateral hypothalamus. This family of peptides modulates sleep-wake cycle, arousal and feeding behaviors; however, the mechanisms regulating their expression remain to be fully elucidated. There is an interest in defining the key molecular elements in orexin regulation, as these may serve to identify targets for generating novel therapies for sleep disorders, obesity and addiction. Our previous studies showed that the expression of orexin was decreased in mice carrying null-mutations of the transcription factor early B-cell factor 2 (ebf2) and that the promoter region of the prepro-orexin (Hcrt) gene contained two putative ebf-binding sites, termed olf-1 sites. In the present study, a minimal promoter region of the murine Hcrt gene was identified, which was able to drive the expression of a luciferase reporter gene in the human 293 cell line. Deletion of the olf1-site proximal to the transcription start site of the Hcrt gene increased reporter gene expression, whereas deletion of the distal olf1-like site decreased its expression. The lentiviral transduction of murine transcription factor ebf2 cDNA into 293 cells increased the gene expression driven by this minimal Hcrt-gene promoter and an electrophoretic mobility shift assays demonstrated that the distal olf1-like sequence was a binding site for ebf2.",
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Sánchez-García, A, Cabral-Pacheco, GA, Zomosa-Signoret, VC, Ortiz-López, R, Camacho, A, Tabera-Tarello, PM, Garnica-López, JA & Vidaltamayo, R 2018, 'Modular organization of a hypocretin gene minimal promoter', Molecular Medicine Reports, vol. 17, n.º 2, pp. 2263-2270. https://doi.org/10.3892/mmr.2017.8142

Modular organization of a hypocretin gene minimal promoter. / Sánchez-García, Adriana; Cabral-Pacheco, Griselda A.; Zomosa-Signoret, Viviana C.; Ortiz-López, Rocío; Camacho, Alberto; Tabera-Tarello, Paulo M.; Garnica-López, José A.; Vidaltamayo, Román.

En: Molecular Medicine Reports, Vol. 17, N.º 2, 01.02.2018, p. 2263-2270.

Resultado de la investigación

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T1 - Modular organization of a hypocretin gene minimal promoter

AU - Sánchez-García, Adriana

AU - Cabral-Pacheco, Griselda A.

AU - Zomosa-Signoret, Viviana C.

AU - Ortiz-López, Rocío

AU - Camacho, Alberto

AU - Tabera-Tarello, Paulo M.

AU - Garnica-López, José A.

AU - Vidaltamayo, Román

PY - 2018/2/1

Y1 - 2018/2/1

N2 - Orexins or hypocretins are neurotransmitters produced by a small population of neurons in the lateral hypothalamus. This family of peptides modulates sleep-wake cycle, arousal and feeding behaviors; however, the mechanisms regulating their expression remain to be fully elucidated. There is an interest in defining the key molecular elements in orexin regulation, as these may serve to identify targets for generating novel therapies for sleep disorders, obesity and addiction. Our previous studies showed that the expression of orexin was decreased in mice carrying null-mutations of the transcription factor early B-cell factor 2 (ebf2) and that the promoter region of the prepro-orexin (Hcrt) gene contained two putative ebf-binding sites, termed olf-1 sites. In the present study, a minimal promoter region of the murine Hcrt gene was identified, which was able to drive the expression of a luciferase reporter gene in the human 293 cell line. Deletion of the olf1-site proximal to the transcription start site of the Hcrt gene increased reporter gene expression, whereas deletion of the distal olf1-like site decreased its expression. The lentiviral transduction of murine transcription factor ebf2 cDNA into 293 cells increased the gene expression driven by this minimal Hcrt-gene promoter and an electrophoretic mobility shift assays demonstrated that the distal olf1-like sequence was a binding site for ebf2.

AB - Orexins or hypocretins are neurotransmitters produced by a small population of neurons in the lateral hypothalamus. This family of peptides modulates sleep-wake cycle, arousal and feeding behaviors; however, the mechanisms regulating their expression remain to be fully elucidated. There is an interest in defining the key molecular elements in orexin regulation, as these may serve to identify targets for generating novel therapies for sleep disorders, obesity and addiction. Our previous studies showed that the expression of orexin was decreased in mice carrying null-mutations of the transcription factor early B-cell factor 2 (ebf2) and that the promoter region of the prepro-orexin (Hcrt) gene contained two putative ebf-binding sites, termed olf-1 sites. In the present study, a minimal promoter region of the murine Hcrt gene was identified, which was able to drive the expression of a luciferase reporter gene in the human 293 cell line. Deletion of the olf1-site proximal to the transcription start site of the Hcrt gene increased reporter gene expression, whereas deletion of the distal olf1-like site decreased its expression. The lentiviral transduction of murine transcription factor ebf2 cDNA into 293 cells increased the gene expression driven by this minimal Hcrt-gene promoter and an electrophoretic mobility shift assays demonstrated that the distal olf1-like sequence was a binding site for ebf2.

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Sánchez-García A, Cabral-Pacheco GA, Zomosa-Signoret VC, Ortiz-López R, Camacho A, Tabera-Tarello PM y otros. Modular organization of a hypocretin gene minimal promoter. Molecular Medicine Reports. 2018 feb 1;17(2):2263-2270. https://doi.org/10.3892/mmr.2017.8142