Gata4 is an important transcription factor in heart development. Gata4 post-transcriptional protein modification regulates transcriptional activity and DNA binding, which in turn affects expression of downstream genes and transcription factors, differentiation of embryonic stem cells and cardiogenesis. This article summarizes the effect of post-transcriptional protein modification on transcriptional activity of Gata4 and the relationship between this effect and congenital heart disease. It was shown that acetylation, phosphorylation and SUMOylation upregulate transcriptional activity, DNA binding, downstream gene expression and embryonic stem cell differentiation. On the other hand, methylation and deacetylation downregulate Gata4 transcriptional activity. Post-transcriptional protein modification of Gata4 is very important in clinical research on congenital and other heart diseases.
Acetylation ; Animals ; GATA4 Transcription Factor ; chemistry ; genetics ; metabolism ; Humans ; Methylation ; Phosphorylation ; Protein Processing, Post-Translational ; Sumoylation

OBJECTIVETo identify the genetic defects in patients with congenital atrial septal defects (ASD).

METHODSThe clinical data and blood samples from 180 unrelated subjects with congenital ASD were collected and evaluated. Two hundred healthy individuals served as controls. The coding exons and the flanking introns of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with the sequences publicized in GenBank by the aid of programme BLAST to identify the sequence variations. Clustal W software was applied for analysis of the conservation of altered amino acids.

RESULTSTwo novel heterozygous missense GATA4 mutations were identified in 2 out of 180 ASD patients. Namely, the triplet substitutions of GTC for GGC at codon 21 and TCG for CCG at codon 87 were detected, predicting the conversions of glycine into valine at amino acid residue 21 (G21V) and proline into serine at amino acid residue 87 (P87S). None of the two mutations were detected in 200 healthy controls. Across-species alignment of GATA4 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, a single nucleotide polymorphism c.99G>T was observed. However, the polymorphic frequency distribution in ASD cases was similar with that in healthy controls (for genotype GT, χ(2) = 0.7556, P = 0.3847; for allele T, χ(2) = 0.7235, P = 0.3950).

CONCLUSIONSTwo novel mutations of GATA4 gene are identified in two unrelated ASD patients. This finding provides new insight into the molecular etiology responsible for ASD.

Case-Control Studies ; Child, Preschool ; DNA Mutational Analysis ; GATA4 Transcription Factor ; genetics ; Genome ; Heart Septal Defects, Atrial ; genetics ; Humans ; Mutation

OBJECTIVE: To explore the correlation between DSG2, TTN and GATA4 genes and Brugada syndrome in Henan Province of China. METHODS: From February 2017 to February 2019, 100 patients with Brugada syndrome and 100 healthy individuals were selected as the study and the control groups, respectively. Electrocardiogram and echocardiography were carried out, and peripheral blood samples was collected. Coding regions of DSG2, TTN and GATA4 genes were amplified by PCR and sequenced. The results were compared with standard sequences from GenBank. RESULTS: Electrocardiogram showed that all patients from the study group had ventricular arrhythmia, 87 cases (87%) presented ventricular tachycardia (VT), 84 cases (84%) presented T wave inversion, and 51 cases (51%) presented Epsilon wave. Echocardiography showed that the right ventricle in the study group was enlarged with the inner diameter of the right ventricle being (40.0±13.3) mm, and the right ventricle showed various degree of abnormal systolic function. The enlargement of right atrium accounted for 64%, and the involvement of the left ventricle accounted for 27%. The right ventricular diameter and left ventricular diastolic diameter of the study group were significantly greater than those of the control group (P< 0.05). DNA sequencing showed that 60 patients carried DSG2 gene variants, among which 18 had missense variant of exon 8. Fifty patients carried TTN gene variants, including 8 in the A-band domain and 3 in the I-band domain. Twenty patients carried 3 variants of the GATA4 gene. CONCLUSION Variants of the DSG2, TTN and GATA4 genes in Henan region are correlated with the onset of Brugada syndrome.
Arrhythmogenic Right Ventricular Dysplasia ; Brugada Syndrome/genetics* ; China ; Connectin ; Desmoglein 2/genetics* ; GATA4 Transcription Factor ; Humans ; Pedigree ; Sequence Analysis, DNA

Bone Morphogenetic Protein 4 ; genetics ; Cell Adhesion Molecules ; genetics ; Extracellular Matrix Proteins ; genetics ; GATA4 Transcription Factor ; genetics ; Heart Septal Defects, Atrial ; genetics ; Heart Septal Defects, Ventricular ; genetics ; Humans

Disorder of Sex Development, 46,XY/genetics* ; GATA4 Transcription Factor/genetics* ; Genetic Testing ; Heart Defects, Congenital ; Humans ; Male ; Molecular Structure ; Mutation/genetics*

OBJECTIVETo screen the gene GATA4 for novel mutations associated with congenital atrial septal defect (ASD).

METHODSThe clinical data and peripheral venous blood specimen from 85 unrelated subjects with congenital ASD were collected and analyzed in contrast to 200 healthy individuals. The coding exons and the exon/intron boundaries of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination procedure. The obtained sequences were aligned with those publicized in GenBank with the help of programme BLAST to identify the sequence variations. The software Clustal W was applied to analysis of the conservation of altered amino acids.

RESULTSThree novel heterozygous missense GATA4 mutations were identified in 3 of 85 ASD patients, respectively. Namely, the triplet substitutions of ATG for GTG at codon 267, GCC for ACC at codon 354, and CAA for CCA at codon 407, predicting the conversions of valine into methionine at amino acid residue 267 (V267M), threonine into alanine at amino acid residue 354 (T354A), and proline into glutamine at amino acid residue 407 (P407Q), were identified. No mutation was detected in 200 healthy controls. A cross-species alignment of GATA4 encoded protein sequences showed that the valine at amino acid residue 267 and proline at amino acid residue 407 were completely conserved evolutionarily.

CONCLUSIONThree novel heterozygous missense GATA4 mutations were identified in patients with congenital ASD, which reveals new molecular etiology responsible for ASD, and contributes to the early prophylaxis and therapy for ASD.

Base Sequence ; Case-Control Studies ; DNA Mutational Analysis ; GATA4 Transcription Factor ; genetics ; Genetic Testing ; Heart Septal Defects, Atrial ; genetics ; Humans ; Mutation, Missense

OBJECTIVETo study the association of single nucleotide polymorphisms (SNPs) of transcription factors (NKX2.5, GATA4, TBX5, and FOG2) with congenital heart disease (CHD) in the Chinese population.

METHODSPubMed, Google Scholar, CNKI, Wanfang Data, and Weipu Data were searched for articles on the association of SNPs of target genes with CHD in the Chinese population. If one locus was mentioned in at least two articles, the random or fixed effect model was used to perform a pooled analysis of study results and to calculate the pooled OR and its 95%CI. If a locus was mentioned in only one article, related data were extracted from this article to analyze the association between the SNPs of this locus and CHD.

RESULTSTwenty-three articles were included. The Meta analysis showed that there were significant differences between the CHD and control groups in the genotype and allele frequencies of GATA4 rs1139244 and rs867858 and the genotype frequency of GATA4 rs904018, while there were no significant differences in the SNPs of the other genetic loci between the two groups. The single-article analysis showed that there were significant differences between the two groups in the allele frequencies of NKX2.5 rs118026695/rs703752, GATA4 rs884662/rs12825/rs12458/rs3203358/rs4841588, and TBX5 rs6489956. There were no significant differences in the SNPs of FOG2 locus between the two groups.

CONCLUSIONSThe SNPs of some loci in NKX2.5, GATA4, and TBX5 are associated with CHD in the Chinese population, but the association between the SNPs of FOG2 locus and the development of CHD has not been found yet.

Asian Continental Ancestry Group ; genetics ; DNA-Binding Proteins ; genetics ; GATA4 Transcription Factor ; genetics ; Genetic Predisposition to Disease ; Heart Defects, Congenital ; genetics ; Homeobox Protein Nkx-2.5 ; genetics ; Humans ; Polymorphism, Single Nucleotide ; T-Box Domain Proteins ; genetics ; Transcription Factors ; genetics

OBJECTIVETo investigate the role of GATA4 gene in the endocardial cushions development.

METHODSTarget gene eukaryote expression vectors were constructed by pcDNA3.1(-) vector plasmid, and were identified by DNA sequence analysis. Recombinant plasmids were transfected into Hela cells with lipofectamine 2000, meanwhile Hela cells transfected with empty vector or those without transfection served as transfection control group and blank control group, respectively. Real-time PCR and Western blot were performed to detect the relative expression of mRNA and protein of transcription factors GATA4, Sox9, Scleraxis and ECM proteins Aggrecan, Tenascin in each group.

RESULTSThe relative mRNA expression of GATA4 in experimental group was significantly higher than in transfection control group and blank control group. GATA4 mRNA expression in Hela(GATA4), Hela(H436Y), Hela(Null) and Hela group was 310.83 ± 2.39, 146.35 ± 1.74, 0.94 ± 0.32, 1.00 ± 0.28, respectively (F = 72.508, P < 0.05). Western blot results were consistent with the results obtained by qRT-PCR. The relative mRNA and protein expressions of Sox9, Scleraxis, Aggrecan and Tenascin in both experimental groups were significantly higher than that in transfection control group and blank control group (P < 0.05), and above gene expressions were significantly downregulated in GATA4(H436Y) group, while they were similar between transfection control group and blank control group (all P > 0.05).

CONCLUSIONSGATA4 H436Y mutation reduces it's transcriptional activation, which might serve as a theoretical framework to demonstrate the roles of GATA4 gene in endocardial cushion development.

Aggrecans ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Down-Regulation ; Endocardial Cushions ; embryology ; GATA4 Transcription Factor ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; HeLa Cells ; Humans ; RNA, Messenger ; SOX9 Transcription Factor ; metabolism ; Tenascin ; metabolism ; Transfection

OBJECTIVETo investigate the effect of cardiotrophin-1 (CT-1) on the GATA4 expression and related signaling pathways (JAK-STAT3, ERK1/2 and PI3-K) in rat cardiomyocytes.

METHODSUsing semi-quantitative RT-PCR and EMSA, we measured the dose and time dependent effects of CT-1 on GATA4 mRNA and binding activity in cultured rat cardiomyocytes. Parthenolide (a STAT inhibitor), U-0126 (an ERK inhibitor) and LY-294002 (a PI3-K inhibitor) alone or in combination were added to the culture medium to assess the role of above signaling pathways in CT-1 mediated effects.

RESULTSGATA4 mRNA expression significantly increased at 3 h post 0.1 nmol/L CT-1 exposure, peaked at 6 h and remained high till 24 h post exposure. The GATA4 binding activity began to increase at 10 min and peaked at 60 min and returned to baseline level 180 min. Six hours post CT-1 (0.01 nmol/L, 0.1 nmol/L, 1 nmol/L) exposure, the GATA4 mRNA expression increased in a dose-dependent manner. The GATA4 binding activity peaked with 0.1 nmol/L CT-1 and higher dose did not further increase the binding activity. U-0126 increased the GATA4 mRNA expression and enhanced the GATA4 binding activity and these effects could be partially attenuated with addition of Parthenolide. Parthenolide also prevented the increase of GATA4 mRNA and binding activity induced by CT-1. LY-294002 had no effects GATA4 mRNA and binding activity.

CONCLUSIONCT-1 increases the GATA4 mRNA expression and binding activity in rat cardiomyocytes via STAT3/ERK1/2 pathways and these effects are independent of PI3-K pathway.

Animals ; Cell Line ; Cytokines ; pharmacology ; GATA4 Transcription Factor ; biosynthesis ; genetics ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; STAT3 Transcription Factor ; pharmacology ; Signal Transduction

BACKGROUNDIn a suitable microenvironment, bone marrow mesenchymal stem cells (BMSCs) can transdifferentiate into myocardial cells whose special gene can be expressed as structural proteins. Growth factor (GF) plays an important role in the cell migration, survival and differentiation. However, the effect of GF on the cellular differentiation is not well understood. In this study, the hepatocyte growth factor (HGF) and insulin like growth factor-1 (IGF-1) were used in the mixed culture of BMSCs and myocardial cells and the effects of these growth factors on the GATA-4 expression of BMSCs were investigated.

METHODSBMSCs were isolated from the marrow of rabbit femurs and tibias and foetal rabbit ventricular myocytes were isolated with trypsin sequential digestion. These two kinds of cells were cocultured in a ratio of 1:1 for 6 weeks; cocultured cells with added HGF and IGF-1 were the experimental group. The differentiated BMSCs were collected using the laser capture, microdissection system and their RNA isolated. Immunocytochemical staining, transmission electron microscopy and reverse transcription-polymerase chain reaction were used to evaluate the transformation of the stem cells into cardiomyocytes like cells.

RESULTSWhen cultured separately, BMSCs did not express alpha-actin and the stem cells had many nucleoli. However, when cocultured with cardiomyocytes, BMSCs expressed alpha-actin and the cardiac transcription factor GATA-4 and showed cardiomyocyte like ultrastructure. In comparison with the control group, the experimental group exhibited the enhanced expression level of GATA-4. The GATA-4 expression of BMSCs increased gradually following the addition of HGF and IGF-1, reached the maximal level after two weeks and decreased slightly thereafter.

CONCLUSIONSBMSCs can transdifferentiate into cardiomyocytes like cells and express the cardiac transcription factor GATA-4 after being cocultured with myocardial cells. HGF and IGF-1 can stimulate transdifferentiation of BMSCs into cardiac phenotype and enhance the expression of GATA-4. These results indicate that growth factors have a great potential in clinical cellular therapy.

Animals ; Cell Differentiation ; drug effects ; Coculture Techniques ; GATA4 Transcription Factor ; genetics ; Hepatocyte Growth Factor ; pharmacology ; Insulin-Like Growth Factor I ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Rabbits