GATA-4 Gene Organization and Analysis of Its Promoter

Abstract

The GATA DNA-binding protein family in mammals has two tandem zinc fingers separated by 29 amino acid residues [(CX 2 C)X 17 (CX 2 C)]-X 29 -[(CX 2 C)X 17 (CX 2 C)] in the middle of the molecule. These fingers are essential for both DNA binding and interaction with other regulatory factors.1,2) The mammalian GATA family has six members. They are divided in two subgroups; one group consists of GATA-1, -2 and -3, which play crucial roles in the development and gene regulation of hematopoietic lineage cells. The other subfamily members, GATA-4, -5 and -6, are similarly important in tissues derived from the endoderm and mesoderm such as the gut, heart and gonad. 1)GATA-4 plays an essential role in embryogenesis, since GATA-4 null mice die at 8.5-10.5 d post-coitum, 3,4) possibly due to the defective ventral body pattern lacking a ventral pericardiac cavity, heart tube and foregut. Since knockout mice exhibited specification of the cardiac cell lineage, 3,4) and ES cells with homozygous GATA-4(Ϫ/Ϫ) contributed to the heart tissue in chimeric embryos, 3) GATA-4 in procardiomyocytes or visceral endoderm cells likely regulate the expression of a morphogenic signaling molecule(s) that directs the migration of procardiomyocytes. Although GATA-4 knockout mice express cardiac contractile proteins, 3,4) a number of studies have suggested that both structural and regulatory genes in the cardiac muscle cells are targets of GATA-4, such as those of A1 adenosine receptor, 5) angiotensin II type Ia receptor, 6) atrial natriuretic peptide, 7) brain natriuretic peptide, 7-9) cardiac a actin, 10) cardiac troponin C, 11) cardiac troponin I, 9) cardiac-restricted ankyrin repeat protein, 12) amyosin heavy-chain, 9,13) b-myosin heavy-chain, 9) myosin light chain 1/3, 14) Na ϩ /Ca 2ϩ exchanger 1, 15,16) Nkx-2.5, 17,18) slow myosin heavy chain, 19) and corin. 20) It is also evident that GATA-4 functions in the regulation of both epithelial cell differentiation and gene expression in the gut 21,22) and gonads [23][24][25][26][27] in addition to the heart.28)The gene regulation by GATA-4 is cooperative, as the interaction of GATA-4 with other regulatory factors such as AP-1, NF-AT3, 34) GATA-6, 9) dHAND, 35) and CBP 36) has been demonstrated. Actually, mutation of GATA-4 prevented the interaction with FOG2 and TBX5, and affected cardiac development. 32,37) Furthermore, it has been reported that the alteration in GATA-4 expression is closely related to pathological conditions such as tumorigenesis [38][39][40] and heart diseases.41) GATA-4 gene activation is also closely associated with hypertrophic responses.6,42,43) However, little is known about the regulation of GATA-4 transcription itself. In this study, we isolated the 5Ј upstream region of the mouse GATA-4 gene, and characterized its sequence in terms of sequence motifs and promoter activity. Furthermore, we showed that a P19.CL6 embryonic carcinoma cell line 44) that differentiates into cardiomyocytes could be a model for regulation of the GATA-4 gene. MATERIALS AND METHODSSequence...

Smooth and Cardiac Muscle-selective Knock-out of Krüppel-like Factor 4 Causes Postnatal Death and Growth Retardation

“…Guittot et al (28) show that the first 5-kb of Gata4 promoter-enhancer sequence was sufficient to direct gene expression in the adult mouse heart, although they did not examine the activity of this fragment during embryonic and postnatal development. Recently, two GC boxes and an E box between Ϫ124 bp and Ϫ36 bp of the mouse Gata4 promoter have been shown to be important for its expression during differentiation of P19.CL6 cells into beating cardiomyocyte-like cells in vitro (29). Results of our present studies showed that Klf4 selectively bound to the Gata4 promoter region in the heart of control mice but not conditional Klf4 knock-out mice, as determined by in vivo ChIP assays.…”

“…Although little is known regarding the transcriptional control of the Gata4 gene, results of several studies showed that multiple GC boxes and an E box within its promoter (Fig. 7A) were important for expression (27)(28)(29). We found a consensus Klf4 binding site at Ϫ125/Ϫ119 bp, in close proximity to four GC boxes and an E box (Fig.…”

An E-Box Element in the Proximal GATA4 Promoter Is Required for Gata4 Expression In Vivo.

“…Regarding gonadal expression, we also previously reported that the same 5-kb fragment controls reporter gene expression specifically in the testes and more precisely in Sertoli cells from embryonic to adult stages [20,21]. A detailed analysis of this 5-kb fragment revealed an evolutionary conserved Ebox element (CACGTG) located near the transcriptional start site and we found this regulatory element to be critical for Gata4 promoter activity in vitro, being bound by USF2 in gonadal cell lines [20,22]. However, the importance of this Ebox motif in vivo is unknown.…”

Interaction of Mat-8 (FXYD-3) with Na+/K+-ATPase in Colorectal Cancer Cells

“…The PCR products were size-separated on an agarose gel (1%) and DNA bands were visualized with ethidium bromide. 13) Total RNA of HeLa and HEK-293 cells 14) was also used. Expression Plasmids for Mouse Mat-8 The expression plasmid for mouse Mat-8, pMat-8-DsRed, 11) was digested with NotI and PspOMI to remove the DsRed moiety.…”