A minigene containing four discrete cis elements recapitulates GATA-1 gene expression in vivo

Kinuko Ohneda, Ritsuko Shimizu, Shigeko Nishimura, Yasushi Muraosa, Satoru Takashi, James Douglas Engel, Masayuki Yamamoto

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Background: The GATA-1 haematopoietic enhancer (G1HE), located between 3.9 and 2.6 kb 5′ to the haematopoietic first exon, is essential for GATA-1 gene transcription in erythroid cells. However, G1HE is not sufficient to confer tissue specificity on the GATA-1 gene in vivo, indicating that additional regulatory sequences are necessary. Results: We demonstrate here that two other upstream promoter elements containing a double GATA motif or two CACCC boxes are also indispensable for reporter gene expression in erythroid cells in the transgenic mouse. The combination of these three cis-acting regions was sufficient for reporter expression in primitive erythroid cells, as demonstrated by linking the elements together into a 659 bp artificial (GdC) minigene. The minigene activated the transcription of a reporter gene from either the endogenous or an exogenous thymidine kinase promoter, retaining cell type-specificity. The addition of a 320 bp fragment in the first intron to the GdC minigene sustained reporter expression in the definitive stage. Moreover, a line of transgenic mouse that expressed GATA-1 cDNA under the control of the complete 979 bp minigene rescued GATA-1 germ line mutant mice from embryonic lethality. Conclusions: A combination of four distinct sequence motifs co-operatively serve as a fundamental functional unit for GATA-1 erythroid transcription in vivo.

Original languageEnglish
Pages (from-to)1243-1254
Number of pages12
JournalGenes to Cells
Volume7
Issue number12
DOIs
Publication statusPublished - 2002 Dec 1
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

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