Arrest in Primitive Erythroid Cell Development Caused by Promoter-specific Disruption of the GATA-1 Gene*
- Satoru Takahashi‡,
- Kou Onodera§,
- Hozumi Motohashi‡,
- Naruyoshi Suwabe‡,
- Norio Hayashi§,
- Nobuaki Yanai¶,
- Yoichi Nabesima‖ and
- Masayuki Yamamoto‡**
- From the ‡Center for Tsukuba Advanced Research Alliance and Institute of Basic Medical Sciences, University of Tsukuba, Tennodai, Tsukuba 305, Japan, the §Department of Biochemistry, School of Medicine and ¶Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980–77, Japan, and the ‖Department of Molecular Genetics, National Institute of Neuroscience, Ogawa-higashi, Kodaira 187, Japan
Abstract
To elucidate the in vivo function of GATA-1 during hematopoiesis, we specifically disrupted the erythroid promoter of the GATA-1 gene in embryonic stem cells and generated germ line chimeras. Male offspring of chimeras bearing the targeted mutation were found to die by 12.5 days post coitus due to severe anemia while heterozygous females displayed characteristics ranging from severe anemia to normal erythropoiesis. When female heterozygotes were crossed with transgenic males carrying a reporter gene, which specifically marks primitive erythroid progenitors, massive accumulation of undifferentiated erythroid cells were observed in the yolk sacs of the GATA-1-mutant embryos, demonstrating that GATA-1 is required for the terminal differentiation of primitive erythroid cells in vivo.
Footnotes
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↵* This work was supported by the Grants-in-Aid from the Ministry of Education, Science and Culture, Japanese Society for Promotion of Sciences, and the Uehara Memorial Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵** To whom correspondence should be addressed: Institute of Basic Medical Sciences, University of Tsukuba, 1–1-1 Tennoudai, Tsukuba 305, Japan. Tel.: 81-298-53-3111; Fax: 81-298-53-6965; E-mail:masiya{at}igaku.md.tsukuba.ac.jp.
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↵1 The abbreviations used are; ES, embryonic stem; CFU-E, colony forming unit-erythroid; CFU-GM, colony forming unit-granulocyte/macrophage; dpc, days post coitus; IE exon or promoter, erythroid-specific first exon or promoter, respectively; IT exon or promoter, testis-specific first exon or promoter, respectively; LacZ, β-galactosidase; neo, neomycin-resistance gene; X-gal, 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside; bp, base pair; IL, interleukin; PCR, polymerase chain reaction; RT-PCR, reverse transcriptase-PCR.
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- Received January 24, 1997.
- Revision received February 17, 1997.
