Functional analysis of the congenital heart disease‑associated GATA4 H436Y mutation in�vitro

Abstract

congenital heart disease (cHd) is the most common type of developmental defect, with high rates of morbidity in infants. The transcription factor GaTa-binding factor 4 (GATA4) has been reported to serve a critical role in embryogenesis and cardiac development. our previous study reported a heterozygous GATA4 c.1306c>T (p.H436Y) mutation in four chinese infants with congenital heart defects. in the present study, functional analysis of the GATA4 H436Y mutation was performed in vitro. The functional effect of GATA4 mutation was compared with GATA4 wild-type using a dual-luciferase reporter assay system and immunofluorescence. electrophoretic mobility-shift assays were performed to explore the binding affinity of the mutated GATA4 to the heart and neural crest derivatives expressed 2 (HAND2) gene. The results revealed that the mutation had no effect on normal nuclear localization, but resulted in diminished GATA-binding affinity to HAND2 and significantly decreased gene transcriptional activation. These results indicated that this GATA4 mutation may not influence cellular localization in transfected cells, but may affect the affinity of the GATA-binding site on HAND2 and decrease transcriptional activity, thus suggesting that the GATA4 mutation may be associated with the pathogenesis of cHd.

In silico analysis of GATA4 variants demonstrates main contribution to congenital heart disease

“… 12 GATA4 has been shown to interact with HAND2 to modulate the transcription of the downstream gene by binding to the conserved GATA-binding sites of the HAND2 promoter. 98 NKX2-5, as a central regulator of many aspects of heart development, interacts with SRF and GATA4 to promote the expression of the cardiac sarcomeric protein gene. 99 Mutations in the ZFPM2 gene, which encodes the FOG2 protein (a transcription regulator of the GATA family members), disrupt the interaction with GATA4 or the nucleosome remodeling and deacetylation (NuRD) complex and, thus, lead to CHD.…”