{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T06:24:45Z","timestamp":1766557485346,"version":"build-2065373602"},"reference-count":44,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T00:00:00Z","timestamp":1631750400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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\u2013pituitary\u2013thyroid axis (HPT axis) by regulating the genes encoding prepro-thyrotropin-releasing hormone (preproTRH) and thyroid-stimulating hormone \u03b2 (TSH\u03b2). 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 TSH\u03b2 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 TSH\u03b2 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.<\/jats:p>","DOI":"10.3390\/ijms221810015","type":"journal-article","created":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T21:38:12Z","timestamp":1631828292000},"page":"10015","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Impairment of the Hypothalamus\u2013Pituitary\u2013Thyroid Axis Caused by Naturally Occurring GATA2 Mutations In Vitro"],"prefix":"10.3390","volume":"22","author":[{"given":"Yuki","family":"Sakai","sequence":"first","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8538-8742","authenticated-orcid":false,"given":"Kenji","family":"Ohba","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"},{"name":"Medical Education Center, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shigekazu","family":"Sasaki","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akio","family":"Matsushita","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroko Misawa","family":"Nakamura","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Go","family":"Kuroda","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daisuke","family":"Tsuriya","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miho","family":"Yamashita","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"},{"name":"International Center, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takafumi","family":"Suda","sequence":"additional","affiliation":[{"name":"Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1210\/er.2013-1087","article-title":"Central regulation of hypothalamic-pituitary-thyroid axis under physiological and pathophysiological conditions","volume":"35","author":"Fekete","year":"2014","journal-title":"Endocr. 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