Cargando…
Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro
The transcription factor GATA2 regulates gene expression in several cells and tissues, including hematopoietic tissues and the central nervous system. Recent studies revealed that loss-of-function mutations in GATA2 are associated with hematological disorders. Our earlier in vitro studies showed tha...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8467656/ https://www.ncbi.nlm.nih.gov/pubmed/34576178 http://dx.doi.org/10.3390/ijms221810015 |
_version_ | 1784573454117240832 |
---|---|
author | Sakai, Yuki Ohba, Kenji Sasaki, Shigekazu Matsushita, Akio Nakamura, Hiroko Misawa Kuroda, Go Tsuriya, Daisuke Yamashita, Miho Suda, Takafumi |
author_facet | Sakai, Yuki Ohba, Kenji Sasaki, Shigekazu Matsushita, Akio Nakamura, Hiroko Misawa Kuroda, Go Tsuriya, Daisuke Yamashita, Miho Suda, Takafumi |
author_sort | Sakai, Yuki |
collection | PubMed |
description | The transcription factor GATA2 regulates gene expression in several cells and tissues, including hematopoietic tissues and the central nervous system. Recent studies revealed that loss-of-function mutations in GATA2 are associated with hematological disorders. Our earlier in vitro studies showed that GATA2 plays an essential role in the hypothalamus–pituitary–thyroid axis (HPT axis) by regulating the genes encoding prepro-thyrotropin-releasing hormone (preproTRH) and thyroid-stimulating hormone β (TSHβ). However, the effect of GATA2 mutants on the transcriptional activity of their promoters remains unelucidated. In this study, we created five human GATA2 mutations (R308P, T354M, R396Q, R398W, and S447R) that were reported to be associated with hematological disorders and analyzed their functional properties, including transactivation potential and DNA-binding capacity toward the preproTRH and the TSHβ promoters. Three mutations (T354M, R396Q, and R398W) within the C-terminal zinc-finger domain reduced the basal GATA2 transcriptional activity on both the preproTRH and the TSHβ promoters with a significant loss of DNA binding affinity. Interestingly, only the R398W mutation reduced the GATA2 protein expression. Subsequent analysis demonstrated that the R398W mutation possibly facilitated the GATA2 degradation process. R308P and S447R mutants exhibited decreased transcriptional activity under protein kinase C compared to the wild-type protein. In conclusion, we demonstrated that naturally occurring GATA2 mutations impair the HPT axis through differential functional mechanisms in vitro. |
format | Online Article Text |
id | pubmed-8467656 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-84676562021-09-27 Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro Sakai, Yuki Ohba, Kenji Sasaki, Shigekazu Matsushita, Akio Nakamura, Hiroko Misawa Kuroda, Go Tsuriya, Daisuke Yamashita, Miho Suda, Takafumi Int J Mol Sci Article The transcription factor GATA2 regulates gene expression in several cells and tissues, including hematopoietic tissues and the central nervous system. Recent studies revealed that loss-of-function mutations in GATA2 are associated with hematological disorders. Our earlier in vitro studies showed that GATA2 plays an essential role in the hypothalamus–pituitary–thyroid axis (HPT axis) by regulating the genes encoding prepro-thyrotropin-releasing hormone (preproTRH) and thyroid-stimulating hormone β (TSHβ). However, the effect of GATA2 mutants on the transcriptional activity of their promoters remains unelucidated. In this study, we created five human GATA2 mutations (R308P, T354M, R396Q, R398W, and S447R) that were reported to be associated with hematological disorders and analyzed their functional properties, including transactivation potential and DNA-binding capacity toward the preproTRH and the TSHβ promoters. Three mutations (T354M, R396Q, and R398W) within the C-terminal zinc-finger domain reduced the basal GATA2 transcriptional activity on both the preproTRH and the TSHβ promoters with a significant loss of DNA binding affinity. Interestingly, only the R398W mutation reduced the GATA2 protein expression. Subsequent analysis demonstrated that the R398W mutation possibly facilitated the GATA2 degradation process. R308P and S447R mutants exhibited decreased transcriptional activity under protein kinase C compared to the wild-type protein. In conclusion, we demonstrated that naturally occurring GATA2 mutations impair the HPT axis through differential functional mechanisms in vitro. MDPI 2021-09-16 /pmc/articles/PMC8467656/ /pubmed/34576178 http://dx.doi.org/10.3390/ijms221810015 Text en © 2021 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 Sakai, Yuki Ohba, Kenji Sasaki, Shigekazu Matsushita, Akio Nakamura, Hiroko Misawa Kuroda, Go Tsuriya, Daisuke Yamashita, Miho Suda, Takafumi Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title | Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title_full | Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title_fullStr | Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title_full_unstemmed | Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title_short | Impairment of the Hypothalamus–Pituitary–Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro |
title_sort | impairment of the hypothalamus–pituitary–thyroid axis caused by naturally occurring gata2 mutations in vitro |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8467656/ https://www.ncbi.nlm.nih.gov/pubmed/34576178 http://dx.doi.org/10.3390/ijms221810015 |
work_keys_str_mv | AT sakaiyuki impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT ohbakenji impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT sasakishigekazu impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT matsushitaakio impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT nakamurahirokomisawa impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT kurodago impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT tsuriyadaisuke impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT yamashitamiho impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro AT sudatakafumi impairmentofthehypothalamuspituitarythyroidaxiscausedbynaturallyoccurringgata2mutationsinvitro |