Cargando…

Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain

Eukaryotic gene expression is controlled at multiple levels, including gene transcription and protein translation initiation. One molecule with key roles in both regulatory mechanisms is methyl CpG binding protein 2 (MeCP2). MECP2 gain- and loss-of-function mutations lead to Rett Syndrome and MECP2...

Descripción completa

Detalles Bibliográficos
Autores principales: Buist, Marjorie, El Tobgy, Nada, Shevkoplyas, Danilo, Genung, Matthew, Sher, Annan Ali, Pejhan, Shervin, Rastegar, Mojgan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105805/
https://www.ncbi.nlm.nih.gov/pubmed/35563748
http://dx.doi.org/10.3390/cells11091442
_version_ 1784708128607043584
author Buist, Marjorie
El Tobgy, Nada
Shevkoplyas, Danilo
Genung, Matthew
Sher, Annan Ali
Pejhan, Shervin
Rastegar, Mojgan
author_facet Buist, Marjorie
El Tobgy, Nada
Shevkoplyas, Danilo
Genung, Matthew
Sher, Annan Ali
Pejhan, Shervin
Rastegar, Mojgan
author_sort Buist, Marjorie
collection PubMed
description Eukaryotic gene expression is controlled at multiple levels, including gene transcription and protein translation initiation. One molecule with key roles in both regulatory mechanisms is methyl CpG binding protein 2 (MeCP2). MECP2 gain- and loss-of-function mutations lead to Rett Syndrome and MECP2 Duplication Syndrome, respectively. To study MECP2 gain-of-function, we generated stably transduced human brain cells using lentiviral vectors for both MECP2E1 and MECP2E2 isoforms. Stable overexpression was confirmed by Western blot and immunofluorescence. We assessed the impact of MeCP2E1-E2 gain-of-function on the MeCP2 homeostasis regulatory network (MECP2E1/E2-BDNF/BDNF-miR-132), mTOR-AKT signaling, ribosome biogenesis, markers of chromatin structure, and protein translation initiation. We observed that combined co-transduction of MeCP2 isoforms led to protein degradation of MeCP2E1. Proteosome inhibition by MG132 treatment recovered MeCP2E1 protein within an hour, suggesting its induced degradation through the proteosome pathway. No significant change was detected for translation initiation factors as a result of MeCP2E1, MeCP2E2, or combined overexpression of both isoforms. In contrast, analysis of human Rett Syndrome brains tissues compared with controls indicated impaired protein translation initiation, suggesting that such mechanisms may have differential sensitivity to MECP2 gain- and loss-of-function. Collectively, our results provide further insight towards the dose-dependent functional role of MeCP2 isoforms in the human brain.
format Online
Article
Text
id pubmed-9105805
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-91058052022-05-14 Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain Buist, Marjorie El Tobgy, Nada Shevkoplyas, Danilo Genung, Matthew Sher, Annan Ali Pejhan, Shervin Rastegar, Mojgan Cells Article Eukaryotic gene expression is controlled at multiple levels, including gene transcription and protein translation initiation. One molecule with key roles in both regulatory mechanisms is methyl CpG binding protein 2 (MeCP2). MECP2 gain- and loss-of-function mutations lead to Rett Syndrome and MECP2 Duplication Syndrome, respectively. To study MECP2 gain-of-function, we generated stably transduced human brain cells using lentiviral vectors for both MECP2E1 and MECP2E2 isoforms. Stable overexpression was confirmed by Western blot and immunofluorescence. We assessed the impact of MeCP2E1-E2 gain-of-function on the MeCP2 homeostasis regulatory network (MECP2E1/E2-BDNF/BDNF-miR-132), mTOR-AKT signaling, ribosome biogenesis, markers of chromatin structure, and protein translation initiation. We observed that combined co-transduction of MeCP2 isoforms led to protein degradation of MeCP2E1. Proteosome inhibition by MG132 treatment recovered MeCP2E1 protein within an hour, suggesting its induced degradation through the proteosome pathway. No significant change was detected for translation initiation factors as a result of MeCP2E1, MeCP2E2, or combined overexpression of both isoforms. In contrast, analysis of human Rett Syndrome brains tissues compared with controls indicated impaired protein translation initiation, suggesting that such mechanisms may have differential sensitivity to MECP2 gain- and loss-of-function. Collectively, our results provide further insight towards the dose-dependent functional role of MeCP2 isoforms in the human brain. MDPI 2022-04-24 /pmc/articles/PMC9105805/ /pubmed/35563748 http://dx.doi.org/10.3390/cells11091442 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Buist, Marjorie
El Tobgy, Nada
Shevkoplyas, Danilo
Genung, Matthew
Sher, Annan Ali
Pejhan, Shervin
Rastegar, Mojgan
Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title_full Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title_fullStr Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title_full_unstemmed Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title_short Differential Sensitivity of the Protein Translation Initiation Machinery and mTOR Signaling to MECP2 Gain- and Loss-of-Function Involves MeCP2 Isoform-Specific Homeostasis in the Brain
title_sort differential sensitivity of the protein translation initiation machinery and mtor signaling to mecp2 gain- and loss-of-function involves mecp2 isoform-specific homeostasis in the brain
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105805/
https://www.ncbi.nlm.nih.gov/pubmed/35563748
http://dx.doi.org/10.3390/cells11091442
work_keys_str_mv AT buistmarjorie differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT eltobgynada differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT shevkoplyasdanilo differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT genungmatthew differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT sherannanali differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT pejhanshervin differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain
AT rastegarmojgan differentialsensitivityoftheproteintranslationinitiationmachineryandmtorsignalingtomecp2gainandlossoffunctioninvolvesmecp2isoformspecifichomeostasisinthebrain