mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity
Autor(a) principal: | |
---|---|
Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.1/18239 |
Resumo: | Forkhead box O (FOXO) proteins are transcription factors involved in cancer and aging and their pharmacological manipulation could be beneficial for the treatment of cancer and healthy aging. FOXO proteins are mainly regulated by post-translational modifications including phosphorylation, acetylation and ubiquitination. As these modifications are reversible, activation and inactivation of FOXO factors is attainable through pharmacological treatment. One major regulatory input of FOXO signaling is mediated by protein kinases. Here, we use specific inhibitors against different kinases including PI3K, mTOR, MEK and ALK, and other receptor tyrosine kinases (RTKs) to determine their effect on FOXO3 activity. While we show that inhibition of PI3K efficiently drives FOXO3 into the cell nucleus, the dual PI3K/mTOR inhibitors dactolisib and PI-103 induce nuclear FOXO translocation more potently than the PI3Kδ inhibitor idelalisib. Furthermore, specific inhibition of mTOR kinase activity affecting both mTORC1 and mTORC2 potently induced nuclear translocation of FOXO3, while rapamycin, which specifically inhibits the mTORC1, failed to affect FOXO3. Interestingly, inhibition of the MAPK pathway had no effect on the localization of FOXO3 and upstream RTK inhibition only weakly induced nuclear FOXO3. We also measured the effect of the test compounds on the phosphorylation status of AKT, FOXO3 and ERK, on FOXO-dependent transcriptional activity and on the subcellular localization of other FOXO isoforms. We conclude that mTORC2 is the most important second layer kinase negatively regulating FOXO activity. |
id |
RCAP_7ee37228992464a407560a4faf1b8f3a |
---|---|
oai_identifier_str |
oai:sapientia.ualg.pt:10400.1/18239 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activityFOXOKinasesmTORHigh content screeningChemical biologyCancer;AgingForkhead box O (FOXO) proteins are transcription factors involved in cancer and aging and their pharmacological manipulation could be beneficial for the treatment of cancer and healthy aging. FOXO proteins are mainly regulated by post-translational modifications including phosphorylation, acetylation and ubiquitination. As these modifications are reversible, activation and inactivation of FOXO factors is attainable through pharmacological treatment. One major regulatory input of FOXO signaling is mediated by protein kinases. Here, we use specific inhibitors against different kinases including PI3K, mTOR, MEK and ALK, and other receptor tyrosine kinases (RTKs) to determine their effect on FOXO3 activity. While we show that inhibition of PI3K efficiently drives FOXO3 into the cell nucleus, the dual PI3K/mTOR inhibitors dactolisib and PI-103 induce nuclear FOXO translocation more potently than the PI3Kδ inhibitor idelalisib. Furthermore, specific inhibition of mTOR kinase activity affecting both mTORC1 and mTORC2 potently induced nuclear translocation of FOXO3, while rapamycin, which specifically inhibits the mTORC1, failed to affect FOXO3. Interestingly, inhibition of the MAPK pathway had no effect on the localization of FOXO3 and upstream RTK inhibition only weakly induced nuclear FOXO3. We also measured the effect of the test compounds on the phosphorylation status of AKT, FOXO3 and ERK, on FOXO-dependent transcriptional activity and on the subcellular localization of other FOXO isoforms. We conclude that mTORC2 is the most important second layer kinase negatively regulating FOXO activity.RTI2018-094629-B-I00MDPISapientiaJimenez, LuciaAmenabar, CarlosMayoral-Varo, VictorMackenzie, Thomas A.Ramos, Maria C.Silva, AndreiaCalissi, GiampaoloGrenho, InêsBlanco-Aparicio, CarmenPastor, JoaquinMegías, DiegoFerreira, BibianaLink, Wolfgang2022-09-09T10:29:04Z2022-08-242022-09-08T13:24:08Z2022-08-24T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.1/18239engMolecules 27 (17): 5414 (2022)1420-304910.3390/molecules27175414info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-06T02:03:24Zoai:sapientia.ualg.pt:10400.1/18239Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:08:03.036392Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
title |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
spellingShingle |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity Jimenez, Lucia FOXO Kinases mTOR High content screening Chemical biology Cancer; Aging |
title_short |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
title_full |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
title_fullStr |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
title_full_unstemmed |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
title_sort |
mTORC2 Is the major second layer kinase negatively regulating FOXO3 activity |
author |
Jimenez, Lucia |
author_facet |
Jimenez, Lucia Amenabar, Carlos Mayoral-Varo, Victor Mackenzie, Thomas A. Ramos, Maria C. Silva, Andreia Calissi, Giampaolo Grenho, Inês Blanco-Aparicio, Carmen Pastor, Joaquin Megías, Diego Ferreira, Bibiana Link, Wolfgang |
author_role |
author |
author2 |
Amenabar, Carlos Mayoral-Varo, Victor Mackenzie, Thomas A. Ramos, Maria C. Silva, Andreia Calissi, Giampaolo Grenho, Inês Blanco-Aparicio, Carmen Pastor, Joaquin Megías, Diego Ferreira, Bibiana Link, Wolfgang |
author2_role |
author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Sapientia |
dc.contributor.author.fl_str_mv |
Jimenez, Lucia Amenabar, Carlos Mayoral-Varo, Victor Mackenzie, Thomas A. Ramos, Maria C. Silva, Andreia Calissi, Giampaolo Grenho, Inês Blanco-Aparicio, Carmen Pastor, Joaquin Megías, Diego Ferreira, Bibiana Link, Wolfgang |
dc.subject.por.fl_str_mv |
FOXO Kinases mTOR High content screening Chemical biology Cancer; Aging |
topic |
FOXO Kinases mTOR High content screening Chemical biology Cancer; Aging |
description |
Forkhead box O (FOXO) proteins are transcription factors involved in cancer and aging and their pharmacological manipulation could be beneficial for the treatment of cancer and healthy aging. FOXO proteins are mainly regulated by post-translational modifications including phosphorylation, acetylation and ubiquitination. As these modifications are reversible, activation and inactivation of FOXO factors is attainable through pharmacological treatment. One major regulatory input of FOXO signaling is mediated by protein kinases. Here, we use specific inhibitors against different kinases including PI3K, mTOR, MEK and ALK, and other receptor tyrosine kinases (RTKs) to determine their effect on FOXO3 activity. While we show that inhibition of PI3K efficiently drives FOXO3 into the cell nucleus, the dual PI3K/mTOR inhibitors dactolisib and PI-103 induce nuclear FOXO translocation more potently than the PI3Kδ inhibitor idelalisib. Furthermore, specific inhibition of mTOR kinase activity affecting both mTORC1 and mTORC2 potently induced nuclear translocation of FOXO3, while rapamycin, which specifically inhibits the mTORC1, failed to affect FOXO3. Interestingly, inhibition of the MAPK pathway had no effect on the localization of FOXO3 and upstream RTK inhibition only weakly induced nuclear FOXO3. We also measured the effect of the test compounds on the phosphorylation status of AKT, FOXO3 and ERK, on FOXO-dependent transcriptional activity and on the subcellular localization of other FOXO isoforms. We conclude that mTORC2 is the most important second layer kinase negatively regulating FOXO activity. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-09-09T10:29:04Z 2022-08-24 2022-09-08T13:24:08Z 2022-08-24T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.1/18239 |
url |
http://hdl.handle.net/10400.1/18239 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Molecules 27 (17): 5414 (2022) 1420-3049 10.3390/molecules27175414 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
MDPI |
publisher.none.fl_str_mv |
MDPI |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799133326419689472 |