Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway

In addition to the well-known function of ACTH as the main regulator of adrenal steroidogenesis, we have previously demonstrated its effect on the transcriptional stimulation of HO-1 expression, a component of the cellular antioxidant defense system. In agreement, we hereby demonstrate that, in adre...

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Autores principales:	Astort, F., Repetto, E.M., Rocha-Viegas, L., Mercau, M.E., Sanchez Puch, S., Finkielstein, C.V., Pecci, A., Cymeryng, C.B.
Formato:	JOUR
Materias:	Adrenocortical cells CAMP/PKA CREB Heme oxygenase PI3K corticotropin cyclic AMP cyclic AMP responsive element binding protein heme oxygenase 1 luciferase n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide phosphatidylinositol 3 kinase protein kinase B reactive oxygen metabolite transcription factor Nrf2 protein binding adrenal cortex cell animal cell Article controlled study enzyme activation enzyme activity enzyme induction enzyme inhibition gene overexpression mouse nonhuman priority journal promoter region protein phosphorylation transcription initiation site transcription regulation transient transfection adrenal gland animal biological model cell line drug effects gene expression regulation genetics metabolism signal transduction Adrenal Glands Adrenocorticotropic Hormone Animals Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Heme Oxygenase-1 Mice Models, Biological Phosphatidylinositol 3-Kinases Promoter Regions, Genetic Protein Binding Reactive Oxygen Species Signal Transduction
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_09525041_v57_n2_p113_Astort
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_09525041_v57_n2_p113_Astort
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spelling	todo:paper_09525041_v57_n2_p113_Astort2023-10-03T15:50:45Z Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway Astort, F. Repetto, E.M. Rocha-Viegas, L. Mercau, M.E. Sanchez Puch, S. Finkielstein, C.V. Pecci, A. Cymeryng, C.B. Adrenocortical cells CAMP/PKA CREB Heme oxygenase PI3K corticotropin cyclic AMP cyclic AMP responsive element binding protein heme oxygenase 1 luciferase n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide phosphatidylinositol 3 kinase protein kinase B reactive oxygen metabolite transcription factor Nrf2 cyclic AMP responsive element binding protein heme oxygenase 1 phosphatidylinositol 3 kinase protein binding reactive oxygen metabolite adrenal cortex cell animal cell Article controlled study enzyme activation enzyme activity enzyme induction enzyme inhibition gene overexpression mouse nonhuman priority journal promoter region protein binding protein phosphorylation transcription initiation site transcription regulation transient transfection adrenal gland animal biological model cell line drug effects gene expression regulation genetics metabolism signal transduction Adrenal Glands Adrenocorticotropic Hormone Animals Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Heme Oxygenase-1 Mice Models, Biological Phosphatidylinositol 3-Kinases Promoter Regions, Genetic Protein Binding Reactive Oxygen Species Signal Transduction In addition to the well-known function of ACTH as the main regulator of adrenal steroidogenesis, we have previously demonstrated its effect on the transcriptional stimulation of HO-1 expression, a component of the cellular antioxidant defense system. In agreement, we hereby demonstrate that, in adrenocortical Y1 cells, HO-1 induction correlates with a significant prevention of the generation of reactive oxygen species induced by H2O2/Fe2+. ACTH/cAMP-dependent activation of redox-imbalanced related factors such as NRF2 or NFκB and the participation of MAPKs in this mechanism was, however, discarded based on results with specific inhibitors and reporter plasmids. We suggest the involvement of CREB in HO-1 induction by ACTH/cAMP, as transfection of cells with a dominant-negative isoform of CREB (DN-CREB-M1) decreased, while overexpression of CREB increased HO-1 protein levels. Sequence screening of the murine HO-1 promoter revealed CRE-like sites located at −146 and −37 of the transcription start site and ChIP studies indicated that this region recruits phosphorylated CREB (pCREB) upon cAMP stimulation in Y1 cells. In agreement, H89 (PKA inhibitor) or cotransfection with DN-CREB-M1 prevented the 8Br-cAMP-dependent increase in luciferase activity in cells transfected with pHO-1[−295/+74].LUC. ACTH and cAMP treatment induced the activation of the PI3K/Akt signaling pathway in a PKA-independent mechanism. Inhibition of this pathway prevented the cAMP-dependent increase in HO-1 protein levels and luciferase activity in cells transfected with pHO-1[−295/+74].LUC. Finally, here we show a crosstalk between the cAMP/PKA and PI3K pathways that affects the binding of p-CREB to its cognate element in the murine promoter of the Hmox1 gene. © 2016 Society for Endocrinology Printed in Great Britain. Fil:Astort, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Rocha-Viegas, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Finkielstein, C.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pecci, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cymeryng, C.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_09525041_v57_n2_p113_Astort
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Adrenocortical cells CAMP/PKA CREB Heme oxygenase PI3K corticotropin cyclic AMP cyclic AMP responsive element binding protein heme oxygenase 1 luciferase n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide phosphatidylinositol 3 kinase protein kinase B reactive oxygen metabolite transcription factor Nrf2 cyclic AMP responsive element binding protein heme oxygenase 1 phosphatidylinositol 3 kinase protein binding reactive oxygen metabolite adrenal cortex cell animal cell Article controlled study enzyme activation enzyme activity enzyme induction enzyme inhibition gene overexpression mouse nonhuman priority journal promoter region protein binding protein phosphorylation transcription initiation site transcription regulation transient transfection adrenal gland animal biological model cell line drug effects gene expression regulation genetics metabolism signal transduction Adrenal Glands Adrenocorticotropic Hormone Animals Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Heme Oxygenase-1 Mice Models, Biological Phosphatidylinositol 3-Kinases Promoter Regions, Genetic Protein Binding Reactive Oxygen Species Signal Transduction
spellingShingle	Adrenocortical cells CAMP/PKA CREB Heme oxygenase PI3K corticotropin cyclic AMP cyclic AMP responsive element binding protein heme oxygenase 1 luciferase n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide phosphatidylinositol 3 kinase protein kinase B reactive oxygen metabolite transcription factor Nrf2 cyclic AMP responsive element binding protein heme oxygenase 1 phosphatidylinositol 3 kinase protein binding reactive oxygen metabolite adrenal cortex cell animal cell Article controlled study enzyme activation enzyme activity enzyme induction enzyme inhibition gene overexpression mouse nonhuman priority journal promoter region protein binding protein phosphorylation transcription initiation site transcription regulation transient transfection adrenal gland animal biological model cell line drug effects gene expression regulation genetics metabolism signal transduction Adrenal Glands Adrenocorticotropic Hormone Animals Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Heme Oxygenase-1 Mice Models, Biological Phosphatidylinositol 3-Kinases Promoter Regions, Genetic Protein Binding Reactive Oxygen Species Signal Transduction Astort, F. Repetto, E.M. Rocha-Viegas, L. Mercau, M.E. Sanchez Puch, S. Finkielstein, C.V. Pecci, A. Cymeryng, C.B. Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
topic_facet	Adrenocortical cells CAMP/PKA CREB Heme oxygenase PI3K corticotropin cyclic AMP cyclic AMP responsive element binding protein heme oxygenase 1 luciferase n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide phosphatidylinositol 3 kinase protein kinase B reactive oxygen metabolite transcription factor Nrf2 cyclic AMP responsive element binding protein heme oxygenase 1 phosphatidylinositol 3 kinase protein binding reactive oxygen metabolite adrenal cortex cell animal cell Article controlled study enzyme activation enzyme activity enzyme induction enzyme inhibition gene overexpression mouse nonhuman priority journal promoter region protein binding protein phosphorylation transcription initiation site transcription regulation transient transfection adrenal gland animal biological model cell line drug effects gene expression regulation genetics metabolism signal transduction Adrenal Glands Adrenocorticotropic Hormone Animals Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Heme Oxygenase-1 Mice Models, Biological Phosphatidylinositol 3-Kinases Promoter Regions, Genetic Protein Binding Reactive Oxygen Species Signal Transduction
description	In addition to the well-known function of ACTH as the main regulator of adrenal steroidogenesis, we have previously demonstrated its effect on the transcriptional stimulation of HO-1 expression, a component of the cellular antioxidant defense system. In agreement, we hereby demonstrate that, in adrenocortical Y1 cells, HO-1 induction correlates with a significant prevention of the generation of reactive oxygen species induced by H2O2/Fe2+. ACTH/cAMP-dependent activation of redox-imbalanced related factors such as NRF2 or NFκB and the participation of MAPKs in this mechanism was, however, discarded based on results with specific inhibitors and reporter plasmids. We suggest the involvement of CREB in HO-1 induction by ACTH/cAMP, as transfection of cells with a dominant-negative isoform of CREB (DN-CREB-M1) decreased, while overexpression of CREB increased HO-1 protein levels. Sequence screening of the murine HO-1 promoter revealed CRE-like sites located at −146 and −37 of the transcription start site and ChIP studies indicated that this region recruits phosphorylated CREB (pCREB) upon cAMP stimulation in Y1 cells. In agreement, H89 (PKA inhibitor) or cotransfection with DN-CREB-M1 prevented the 8Br-cAMP-dependent increase in luciferase activity in cells transfected with pHO-1[−295/+74].LUC. ACTH and cAMP treatment induced the activation of the PI3K/Akt signaling pathway in a PKA-independent mechanism. Inhibition of this pathway prevented the cAMP-dependent increase in HO-1 protein levels and luciferase activity in cells transfected with pHO-1[−295/+74].LUC. Finally, here we show a crosstalk between the cAMP/PKA and PI3K pathways that affects the binding of p-CREB to its cognate element in the murine promoter of the Hmox1 gene. © 2016 Society for Endocrinology Printed in Great Britain.
format	JOUR
author	Astort, F. Repetto, E.M. Rocha-Viegas, L. Mercau, M.E. Sanchez Puch, S. Finkielstein, C.V. Pecci, A. Cymeryng, C.B.
author_facet	Astort, F. Repetto, E.M. Rocha-Viegas, L. Mercau, M.E. Sanchez Puch, S. Finkielstein, C.V. Pecci, A. Cymeryng, C.B.
author_sort	Astort, F.
title	Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
title_short	Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
title_full	Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
title_fullStr	Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
title_full_unstemmed	Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway
title_sort	role of creb on heme oxygenase-1 induction in adrenal cells: involvement of the pi3k pathway
url	http://hdl.handle.net/20.500.12110/paper_09525041_v57_n2_p113_Astort
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Role of CREB on heme oxygenase-1 induction in adrenal cells: Involvement of the PI3K pathway

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