Erythroid transcription factor GATA-1 is abundantly transcribed in mouse testis

Abstract

The transcription factor GATA-1 is a fundamental regulator of genes in haematopoietic cell lineages and belongs to a family of factors that bind to the consensus sequence WGATAR. The GATA motif was originally identified in cis-regulatory regions of globin and other erythroid-specific genes, but the range of genes controlled by the GATA factors has since expanded. Members of the GATA transcription factor family share a conserved zinc-finger DNA-binding domain, but the expression profile of each GATA factor is distinct. Here we show that a testis form of murine (m)GATA-1 messenger RNA is transcribed from a promoter located 5' to the erythroid first exon, and the remaining exons (which encode the mGATA-1 protein) are used in common by both testis and erythroid transcripts. We use an anti-mGATA-1 monoclonal antibody to show that the factor expressed in erythroid cells is the same as that found in the seminiferous tubules of murine testis. The GATA-1-expressing cells in 10-week-old testis were found only in contact with the basement membrane of seminiferous tubules, suggesting that GATA-1 regulates genes during the earliest stages of spermatogenesis.

Differential roles of GATA‐1 and GATA‐2 in growth and differentiation of mast cells

“…The properties of the anti-mGATA-1 N6 monoclonal antibody raised in rats have been described previously (Ito et al 1993). The anti-cGATA-2 monoclonal antibody was prepared using bacterially expressed GST/cGATA-2 fusion protein (Ko & Engel 1993).…”

Human small Maf proteins form heterodimers with CNC family transcription factors and recognize the NF-E2 motif

“…Screening was performed using a mouse mafK cDNA fragment as a probe under non-stringent conditions. Duplicate phage lift ®lters (Colony/Plaque Screeen, Du Pont) were made from each plate and processed for hybridization to the probe as described previously (Ito et al, 1993). This was carried out in the presence of 66SSC (16SSC is 0.15 M NaCl plus 0.015 M sodium citrate) and 0.25% of milk at 508C overnight.…”

“…Prokaryotic expression plasmids for Nrf1 and Nrf2 were constructed as follows. First, single strand cDNA was synthesized using RNA isolated from K562 cells and random primers as described earlier (Ito et al, 1993). cDNAs encoding bZip regions of Nrf1 and Nrf2 were then generated by PCR using the single strand cDNA and following primers.…”

“…Aliquots of the RNA samples (2 mg) were electrophoresed on 1% agarose/formaldehyde gels and transferred to nylon ®lters as described previously (Ito et al, 1993). Northern blots containing multiple human tissue RNA samples were purchased from CLONTECH; (each tissue sample contains 2 mg of poly(A) + RNA).…”

Alternative promoters and polyadenylation regulate tissue‐specific expression of Hemogen isoforms during hematopoiesis and spermatogenesis

“…Because GATA-1 is a key regulator of erythroid differentiation and controls transcription from several erythroid genes, it is likely that HEMGN is a downstream target of GATA-1. Of interest, GATA-1 is also expressed as distinct mRNA isoforms in blood cells and testis under the control of two alternative promoters (Ito et al, 1993). While GATA-1 expression in hematopoietic tissues is under the control of the combination of the GATA-1 site and three other distinct regulatory elements (Tsai et al, 1991;Ohneda et al, 2002), the regulatory mechanisms for GATA-1 expression in testis remains to be defined.…”