The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima
The hypoxia-inducible factor (HIF) is a heterodimeric transcription factor composed of a constitutively expressed HIF-β subunit and an oxygen-regulated HIF-α subunit. We have previously defined a hypoxia-inducible transcriptional response in Drosophila melanogaster that is homologous to the mammalia...
Guardado en:
| Autores principales: | , , , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00219533_v118_n23_p5431_Dekanty |
| Aporte de: |
| id |
todo:paper_00219533_v118_n23_p5431_Dekanty |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_00219533_v118_n23_p5431_Dekanty2023-10-03T14:23:29Z The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima Dekanty, A. Lavista-Llanos, S. Irisarri, M. Oldham, S. Wappner, P. Drosphila Hypoxia-inducible factor (HIF) Nuclear localization PI3K pathway Sima hypoxia inducible factor 1alpha hypoxia inducible factor 1beta insulin messenger RNA phosphatidylinositol 3 kinase phosphatidylinositol 3,4,5 trisphosphate 3 phosphatase protein kinase B protein similar protein tango transcription factor transcription factor PAS unclassified drug article cellular distribution Drosophila embryo embryo development enzyme activation gene overexpression immunofluorescence in vivo study nonhuman Northern blotting polyacrylamide gel electrophoresis priority journal protein localization protein stability reverse transcription polymerase chain reaction RNA interference signal transduction transcription regulation upregulation 1-Phosphatidylinositol 3-Kinase Active Transport, Cell Nucleus Animals Aryl Hydrocarbon Receptor Nuclear Translocator Cell Hypoxia Cell Line Cell Nucleus DNA-Binding Proteins Dose-Response Relationship, Drug Drosophila Drosophila Proteins Embryo, Nonmammalian Gene Expression Regulation Hypoxia-Inducible Factor 1, alpha Subunit Insulin Oxygen Proto-Oncogene Proteins c-akt RNA RNA Interference Signal Transduction Transcription, Genetic Drosophila melanogaster Mammalia The hypoxia-inducible factor (HIF) is a heterodimeric transcription factor composed of a constitutively expressed HIF-β subunit and an oxygen-regulated HIF-α subunit. We have previously defined a hypoxia-inducible transcriptional response in Drosophila melanogaster that is homologous to the mammalian HIF-dependent response. In Drosophila, the bHLH-PAS proteins Similar (Sima) and Tango (Tgo) are the functional homologues of the mammalian HIF-α and HIF-β subunits, respectively. HIF-α/Sima is regulated by oxygen at several different levels that include protein stability and subcellular localization. We show here for the first time that insulin can activate HIF-dependent transcription, both in Drosophila S2 cells and in living Drosophila embryos. Using a pharmacological approach as well as RNA interference, we determined that the effect of insulin on HIF-dependent transcriptional induction is mediated by PI3K-AKT and TOR pathways. We demonstrate that stimulation of the transcriptional response involves upregulation of Sima protein but not sima mRNA. Finally, we have analyzed in vivo the effect of the activation of the PI3K-AKT pathway on the subcellular localization of Sima protein. Overexpression of dAKT and dPDK1 in normoxic embryos provoked a major increase in Sima nuclear localization, mimicking the effect of a hypoxic treatment. A similar increase in Sima nuclear localization was observed in dPTEN homozygous mutant embryos, confirming that activation of the PI3K-AKT pathway promotes nuclear accumulation of Sima protein. We conclude that regulation of HIF-α/Sima by the PI3K-AKT-TOR pathway is a major conserved mode of regulation of the HIF-dependent transcriptional response in Drosophila. Fil:Dekanty, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Lavista-Llanos, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Irisarri, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Wappner, P. 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_00219533_v118_n23_p5431_Dekanty |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Drosphila Hypoxia-inducible factor (HIF) Nuclear localization PI3K pathway Sima hypoxia inducible factor 1alpha hypoxia inducible factor 1beta insulin messenger RNA phosphatidylinositol 3 kinase phosphatidylinositol 3,4,5 trisphosphate 3 phosphatase protein kinase B protein similar protein tango transcription factor transcription factor PAS unclassified drug article cellular distribution Drosophila embryo embryo development enzyme activation gene overexpression immunofluorescence in vivo study nonhuman Northern blotting polyacrylamide gel electrophoresis priority journal protein localization protein stability reverse transcription polymerase chain reaction RNA interference signal transduction transcription regulation upregulation 1-Phosphatidylinositol 3-Kinase Active Transport, Cell Nucleus Animals Aryl Hydrocarbon Receptor Nuclear Translocator Cell Hypoxia Cell Line Cell Nucleus DNA-Binding Proteins Dose-Response Relationship, Drug Drosophila Drosophila Proteins Embryo, Nonmammalian Gene Expression Regulation Hypoxia-Inducible Factor 1, alpha Subunit Insulin Oxygen Proto-Oncogene Proteins c-akt RNA RNA Interference Signal Transduction Transcription, Genetic Drosophila melanogaster Mammalia |
| spellingShingle |
Drosphila Hypoxia-inducible factor (HIF) Nuclear localization PI3K pathway Sima hypoxia inducible factor 1alpha hypoxia inducible factor 1beta insulin messenger RNA phosphatidylinositol 3 kinase phosphatidylinositol 3,4,5 trisphosphate 3 phosphatase protein kinase B protein similar protein tango transcription factor transcription factor PAS unclassified drug article cellular distribution Drosophila embryo embryo development enzyme activation gene overexpression immunofluorescence in vivo study nonhuman Northern blotting polyacrylamide gel electrophoresis priority journal protein localization protein stability reverse transcription polymerase chain reaction RNA interference signal transduction transcription regulation upregulation 1-Phosphatidylinositol 3-Kinase Active Transport, Cell Nucleus Animals Aryl Hydrocarbon Receptor Nuclear Translocator Cell Hypoxia Cell Line Cell Nucleus DNA-Binding Proteins Dose-Response Relationship, Drug Drosophila Drosophila Proteins Embryo, Nonmammalian Gene Expression Regulation Hypoxia-Inducible Factor 1, alpha Subunit Insulin Oxygen Proto-Oncogene Proteins c-akt RNA RNA Interference Signal Transduction Transcription, Genetic Drosophila melanogaster Mammalia Dekanty, A. Lavista-Llanos, S. Irisarri, M. Oldham, S. Wappner, P. The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| topic_facet |
Drosphila Hypoxia-inducible factor (HIF) Nuclear localization PI3K pathway Sima hypoxia inducible factor 1alpha hypoxia inducible factor 1beta insulin messenger RNA phosphatidylinositol 3 kinase phosphatidylinositol 3,4,5 trisphosphate 3 phosphatase protein kinase B protein similar protein tango transcription factor transcription factor PAS unclassified drug article cellular distribution Drosophila embryo embryo development enzyme activation gene overexpression immunofluorescence in vivo study nonhuman Northern blotting polyacrylamide gel electrophoresis priority journal protein localization protein stability reverse transcription polymerase chain reaction RNA interference signal transduction transcription regulation upregulation 1-Phosphatidylinositol 3-Kinase Active Transport, Cell Nucleus Animals Aryl Hydrocarbon Receptor Nuclear Translocator Cell Hypoxia Cell Line Cell Nucleus DNA-Binding Proteins Dose-Response Relationship, Drug Drosophila Drosophila Proteins Embryo, Nonmammalian Gene Expression Regulation Hypoxia-Inducible Factor 1, alpha Subunit Insulin Oxygen Proto-Oncogene Proteins c-akt RNA RNA Interference Signal Transduction Transcription, Genetic Drosophila melanogaster Mammalia |
| description |
The hypoxia-inducible factor (HIF) is a heterodimeric transcription factor composed of a constitutively expressed HIF-β subunit and an oxygen-regulated HIF-α subunit. We have previously defined a hypoxia-inducible transcriptional response in Drosophila melanogaster that is homologous to the mammalian HIF-dependent response. In Drosophila, the bHLH-PAS proteins Similar (Sima) and Tango (Tgo) are the functional homologues of the mammalian HIF-α and HIF-β subunits, respectively. HIF-α/Sima is regulated by oxygen at several different levels that include protein stability and subcellular localization. We show here for the first time that insulin can activate HIF-dependent transcription, both in Drosophila S2 cells and in living Drosophila embryos. Using a pharmacological approach as well as RNA interference, we determined that the effect of insulin on HIF-dependent transcriptional induction is mediated by PI3K-AKT and TOR pathways. We demonstrate that stimulation of the transcriptional response involves upregulation of Sima protein but not sima mRNA. Finally, we have analyzed in vivo the effect of the activation of the PI3K-AKT pathway on the subcellular localization of Sima protein. Overexpression of dAKT and dPDK1 in normoxic embryos provoked a major increase in Sima nuclear localization, mimicking the effect of a hypoxic treatment. A similar increase in Sima nuclear localization was observed in dPTEN homozygous mutant embryos, confirming that activation of the PI3K-AKT pathway promotes nuclear accumulation of Sima protein. We conclude that regulation of HIF-α/Sima by the PI3K-AKT-TOR pathway is a major conserved mode of regulation of the HIF-dependent transcriptional response in Drosophila. |
| format |
JOUR |
| author |
Dekanty, A. Lavista-Llanos, S. Irisarri, M. Oldham, S. Wappner, P. |
| author_facet |
Dekanty, A. Lavista-Llanos, S. Irisarri, M. Oldham, S. Wappner, P. |
| author_sort |
Dekanty, A. |
| title |
The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| title_short |
The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| title_full |
The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| title_fullStr |
The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| title_full_unstemmed |
The insulin-PI3K/TOR pathway induces a HIF-dependent transcriptional response in Drosophila by promoting nuclear localization of HIF-α /Sima |
| title_sort |
insulin-pi3k/tor pathway induces a hif-dependent transcriptional response in drosophila by promoting nuclear localization of hif-α /sima |
| url |
http://hdl.handle.net/20.500.12110/paper_00219533_v118_n23_p5431_Dekanty |
| work_keys_str_mv |
AT dekantya theinsulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT lavistallanoss theinsulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT irisarrim theinsulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT oldhams theinsulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT wappnerp theinsulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT dekantya insulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT lavistallanoss insulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT irisarrim insulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT oldhams insulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima AT wappnerp insulinpi3ktorpathwayinducesahifdependenttranscriptionalresponseindrosophilabypromotingnuclearlocalizationofhifasima |
| _version_ |
1782024676648681472 |