Preliminary functional analysis of a novel mutation in GATA-4 gene in Chinese patients with congenital cardiac septal defects.
- Author:
Ming-wu CHEN
1
;
Yu-sheng PANG
;
Ying GUO
;
Bing-li LIU
;
Jie SHEN
;
Huai-dong SONG
;
Tang-wei LIU
Author Information
- Publication Type:Journal Article
- MeSH: Adolescent; Adult; Child; Child, Preschool; Female; GATA4 Transcription Factor; genetics; Heart Septal Defects; genetics; Humans; Infant; Male; Molecular Sequence Data; Mutagenesis, Site-Directed; Sequence Alignment; Sequence Analysis, DNA; Young Adult
- From: Chinese Journal of Medical Genetics 2009;26(3):277-281
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo perform the functional analysis of a novel H436Y mutation of GATA-4 gene identified in Han Chinese patients with congenital cardiac septal defects.
METHODSUsing bioinformatics to predict if the H436Y mutation in the GATA-4 gene affects its protein function. H436Y mutation in the GATA-4 gene was generated by Quick Change Lightning site-directed mutagenesis kit and verified by DNA sequencing. GATA-4-wt or GATA-4-mut DNA was cotransfected into Hela cells with DNA for the luciferase reporter gene atrial natriuretic factor (ANF), and luciferase activity was measured by an LKB luminometer 48 h after transient transfection.
RESULTSAlignment of the GATA-4 amino acid sequence indicated that the histidine residue at position 436 was conserved, and H436Y mutation in the GATA-4 gene is expected to affect its protein function. The H436Y mutation significantly reduced the transcriptional activation of downstream reporter ANF when compared to wild-type GATA-4 (P<0.01).
CONCLUSIONThe mutation c.1306C-->T of the GATA-4 gene impaired the activation of the downstream target, suggesting that the H436Y mutation in the C-terminal region of the GATA-4 gene might prevent its biological function.