The two zinc finger-like domains of GATA-1 have different DNA binding specificities.

Whyatt, David; deBoer, E.; Grosveld, Frank

doi:10.1002/j.1460-2075.1993.tb06193.x

Cited by 113 publications

(98 citation statements)

References 61 publications

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“…Using both sets of initial random DNA oligomers, greater than 75% of the clones sequenced show that an AGATAA binding sequence is selected, followed by CGATAA (11%) and AGATAG (8%). This strong preference for a GATA motif reflects the binding preferences noted for the full-length cGATA-1 (13) and the C-terminal zinc finger (4,14) and contrasts the reported GATC specificities of the N-terminal zinc finger (9,14) suggesting a role for the C-terminal arm of the cGATA-1 DNA binding domain. It would have been instructive to carry out binding site selection experiments with the Nterminal finger of GATA-1, thus allowing for a direct comparison with the data in Table I.…”

Section: Resultsmentioning

confidence: 75%

Determinants of GATA-1 Binding to DNA

Ghirlando

Trainor

2003

Journal of Biological Chemistry

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Mammalian GATA transcription factors are expressed in various tissues in a temporally regulated manner. The prototypic member, GATA-1, is required for normal erythroid, megakaryocytic, and mast cell development. This family of DNA-binding proteins recognizes a consensus (A/T)GATA(A/G) motif and possesses homologous DNA binding domains consisting of two zinc fingers. The C-terminal finger of GATA-1 recognizes the consensus motif with nanomolar affinities, whereas the N-terminal finger shows a binding preference for a GATC motif, albeit with much reduced affinity (K d Ϸ M). The Nterminal finger of GATA-2 also shows a preference for an AGATCT binding site, with an increased affinity attributed to N-and C-terminal flanking basic residues (K d Ϸ nM). To understand the differences in the binding specificities of the N-and C-terminal zinc fingers of GATA-1, we have constructed a series of swapped domain peptides. We show that the specificity for AGATAA over AGATCT arises from the C-terminal non-finger basic domain. Thus, the N-terminal finger binds preferentially to AGATAA once appended to the C-terminal arm of the C-terminal finger. We further show that this specificity arises from the highly conserved QTRNRK residues. The converse is, however, untrue in the case of the C-terminal finger; swapping of QTRNRK with the corresponding LVSKRA does not switch the DNA binding specificity from AGATAA to AGATCT. These results highlight the important role of residues adjacent to the CXXCX 17 CNAC zinc finger motif (i.e. non-finger residues) in the specific recognition of DNA residues.DNA binding zinc fingers of the form CXXCX 17 CNAC characterize the GATA family of transcription factors. GATA-1, the original member of the family possesses two such zinc fingers (1, 2). The C-terminal finger and flanking basic arm are both necessary and sufficient for binding to the GATA recognition sequence, (A/T)GATA(A/G) (3, 4). Even though the single Nterminal finger does not bind DNA independently with high affinity, it does stabilize GATA-1 interactions with some sites crucial for gene expression (3,(5)(6)(7)(8). Recent reports, however, indicate that the N-terminal finger binds DNA with an affinity much lower than that observed for the C-terminal finger with binding occurring preferentially to GATC motifs (9). Despite the similarities between the C-and N-terminal fingers (Fig. 1, peptides CC11 and NN4, respectively), the N-terminal finger does not recognize the AGATAA consensus motif, indicating that the flanking or poorly conserved linker residues may play a key role in DNA recognition. A role for the flanking residues has been pointed out in the case of the N-terminal finger of GATA-2 (10). Like the N-terminal finger of GATA-1, this homologous finger shows a binding preference for GATC motifs. However, because of the additional basic region at the N terminus a higher binding affinity has been reported for this finger.Solution NMR studies of the complex formed between the C-terminal finger of cGATA-1 and the AGATAA DNA sequence highlight t...

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Section: Resultsmentioning

confidence: 75%

Determinants of GATA-1 Binding to DNA

Ghirlando

Trainor

2003

Journal of Biological Chemistry

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“…In addition, the non-canonical GATA-1 binding sequence, GATT that overlaps with the distal CCAAT motif of the KLF13 gene promoter (Fig. 4), is supposedly recognized by the GATA-1 carboxyl finger [18]. Therefore, the binding of GATA-1 to the promoter region in vivo may not necessarily be ruled out, even though we failed to demonstrate it in vitro.…”

Section: Discussionmentioning

confidence: 78%

Transcriptional regulation of FKLF-2 (KLF13) gene in erythroid cells

Mitsuma¹,

Asano²,

Kinoshita³

et al. 2005

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expres

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FKLF-2 (KLF13) was cloned from fetal globin-expressing tissues and has been shown to be abundantly expressed in erythroid cells. In this study we examined the transcriptional regulation of the KLF13 gene. A 5.5 kb 5′ flanking region cloned from mouse erythroleukemia (MEL) cell genomic DNA showed that major cis regulatory activities exist in the 550 bp sequence to the unique transcription start site, and that the promoter is more active in K562 cells than in COS-7 cells. The promoter was trans-activated by co-expressed GATA-1 through the sequence containing two CCAAT motifs, suggesting that GATA-1 is involved in the abundant expression of KLF13 mRNA in the erythroid tissue. Dual action, i.e. activating effect in COS-7 and repressive effect in K562 cell, was observed on its own promoter, suggesting a feedback mechanism for the transcriptional control of the KLF13 gene in the erythroid environment. These findings provide an insight on the mechanism of inducible mRNA expression of the KLF13 gene in erythroid cells.

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“…GATA4 belongs to the family of GATA transcription factors that regulate the expression of many genes interacting with the consensus sequence WGATAR through a DNA binding domain that consists of two zinc finger motifs (Whyatt et al 1993). The GATA family consists of 6 isoforms: GATA1, 2, 3…”

Section: Figure 2 Cardiogenic Pathwaysmentioning

confidence: 99%