DNA-binding specificities of the GATA transcription factor family.

Abstract

Members of the GATA family of transcription factors, which are related by a high degree of amino acid sequence identity within their zinc finger DNA-binding domains, each show distinct but overlapping patterns of tissue-restricted expression. Although GATA-1, -2, and -3 have been shown to recognize a consensus sequence derived from regulatory elements in erythroid cell-specific genes, WGATAR (in which W indicates A/T and R indicates A/G), the potential for more subtle differences in the binding preferences of each factor has not been previously addressed. By employing a binding selection and polymerase chain reaction amplification scheme with randomized oligonucleotides, we have determined the binding-site specificities of bacterially expressed chicken GATA-1, -2, and -3 transcription factors. Whereas all three GATA factors bind an AGATAA erythroid consensus motif with high affinity, a second, alternative consensus DNA sequence, AGATCTTA, is also recognized well by GATA-2 and GATA-3 but only poorly by GATA-1. These studies suggest that all three GATA factors are capable of mediating transcriptional effects via a common erythroid consensus DNA-binding motif. Furthermore, GATA-2 and GATA-3, because of their distinct expression patterns and broader DNA recognition properties, may be involved in additional regulatory processes beyond those of GATA-1. The definition of an alternative GATA-2-GATA-3 consensus sequence may facilitate the identification of new target genes in the further elucidation of the roles that these transcription factors play during development.Many transcription factors have been found to be members of highly related multifactor families, and thus their specificity of action must be addressed in order to ascertain their respective functions. Consequently, it is critical to determine which factor, from an array of factors with closely related DNA-binding motifs, acts upon a particular recognition site from a variety of sites with similar sequences. Equally important is the elucidation of the means by which this discrimination is achieved; a variety of mechanisms by which related transcription factors are targeted to distinct regulatory elements have been discovered. Differences in DNA-binding properties direct the zinc finger estrogen and progesterone receptors to their appropriate targets (7,44), as is the case with the Antennapedia-related homeodomain proteins Ultrabithorax and Deforned (8,9). The related retinoic acid, thyroid hormone, and vitamin D receptors exemplify a unique solution to the problem of differential target recognition by discriminating between the spacing and orientation of closely related binding sites of these obligate dimers (27,45

The GATA-4 transcription factor transactivates the cardiac muscle-specific troponin C promoter-enhancer in nonmuscle cells.

“…The first of these sites, termed CEF-1, contains the nucleotide sequence AGATIA, which conforms to one of the high-affinity GATA-binding sites identified by random site selection analyses but differs from the consensus GATA-binding sequence (WGATAR) by a single nucleotide substitution (Fig. 1A) (32,42,51). In addition, the second nuclear protein-binding site in the cTnC enhancer, CEF-2, contains the nucleotide sequence GGATAT, which contains the GATA core motif but differs from the consensus GATA-binding sequence in its flanking residues (Fig.…”

“…For example, basic helix-loop-helix myogenic transcription factors are not expressed in developing heart or precardiac mesoderm (49,63,64), and mice contain-Previous studies have demonstrated that the known GATA transcription factors bind the consensus motif (A/ T)GATA(A/G) (16,73). Random site selection has suggested that additional nonconsensus sequences can bind GATA proteins with high affinity (32,42). Two of the GATA transcription factors (GATA-1 and GATA-3) are expressed in a tissuerestricted fashion and have been shown previously to regulate lineage-specific gene expression in erythroid cells and T lymphocytes, respectively (24,29,33,72,78).…”

Pluripotent stem cells reveal erythroid-specific activities of the GATA1 N-terminus

“…GATA1 binds DNA directly via subdomains of its 2 zinc fingers, most importantly the C-finger (52,53). However, several disease-associated amino acid substitutions within the N-finger inhibit GATA1 binding to target genes in vivo, yet have no effect on in vitro binding to naked DNA templates.…”

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

“…The anti-cGATA-2 monoclonal antibody was prepared using bacterially expressed GST/cGATA-2 fusion protein (Ko & Engel 1993). Briefly, Ϸ 100 mg of GST/cGATA-2 was injected into mice using Titremax adjuvant.…”