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

Fang,Tao; Zhu,Yanjie; Xu,Anlan; Zhang,Yanli; Wu,Qingfa; Huang,Guoying; Sheng,Wei; Chen,Mingwu

doi:10.3892/mmr.2019.10481

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

Authors:
- Tao Fang
- Yanjie Zhu
- Anlan Xu
- Yanli Zhang
- Qingfa Wu
- Guoying Huang
- Wei Sheng
- Mingwu Chen
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Affiliations: Division of Life Sciences and Medicine, The First Affiliated Hospital of University of Science and Technology of China, Hefei, Anhui 230036, P.R. China, Cardiovascular Center, Children's Hospital of Fudan University, Shanghai 201102, P.R. China, Department of Neonatology, Anhui Women and Child Health Care Hospital, Hefei, Anhui 230027, P.R. China, Department of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, P.R. China
Published online on: July 9, 2019 https://doi.org/10.3892/mmr.2019.10481
Pages: 2325-2331
Copyright: © Fang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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.

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