Chen J, Knowles H J, Hebert J L, Hackett B P
The Edward Mallinckrodt Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Clin Invest. 1998 Sep 15;102(6):1077-82. doi: 10.1172/JCI4786.
Winged helix transcription factors play important roles in cellular differentiation and cell-specific gene expression. To define the role of the winged helix factor hepatocyte nuclear factor/forkhead homologue (HFH)-4, a targeted mutation was created in the mouse hfh-4 gene. No expression of HFH-4 was detected in hfh-4(-)/- mice by RNA blot analysis, in situ hybridization, or RT-PCR. hfh-4(-)/- mice were noted to have abnormalities of organ situs consistent with random determination of left-right asymmetry. In addition, a complete absence of cilia was noted in hfh-4(-)/- mice. The hfh-4 gene is thus essential for nonrandom determination of left-right asymmetry and development of ciliated cells. Homozygous mutant mice also exhibited prenatal and postnatal growth failure, perinatal lethality and, in some cases, hydrocephalus. RT-PCR revealed an absence of left-right dynein (lrd) expression in the embryonic lungs of hfh-4(-)/- mice, suggesting that HFH-4 may act by regulating expression of members of the dynein family of genes. The abnormalities in ciliary development and organ situs in hfh-4(-)/- mice are similar to those observed in human congenital syndromes such as Kartagener syndrome. Targeted mutation of hfh-4 thus provides a model for elucidating the mechanisms regulating ciliary development and determination of left-right asymmetry.
翼状螺旋转录因子在细胞分化和细胞特异性基因表达中发挥着重要作用。为了确定翼状螺旋因子肝细胞核因子/叉头同源物(HFH)-4的作用,在小鼠hfh-4基因中产生了靶向突变。通过RNA印迹分析、原位杂交或逆转录聚合酶链反应(RT-PCR),在hfh-4(-)/-小鼠中未检测到HFH-4的表达。注意到hfh-4(-)/-小鼠存在器官位置异常,这与左右不对称的随机确定一致。此外,在hfh-4(-)/-小鼠中发现完全没有纤毛。因此,hfh-4基因对于左右不对称的非随机确定和纤毛细胞的发育至关重要。纯合突变小鼠还表现出产前和产后生长发育迟缓、围产期致死率,在某些情况下还出现脑积水。RT-PCR显示hfh-4(-)/-小鼠胚胎肺中缺乏左右动力蛋白(lrd)表达,这表明HFH-4可能通过调节动力蛋白基因家族成员的表达来发挥作用。hfh-4(-)/-小鼠纤毛发育和器官位置的异常与在人类先天性综合征如卡塔格内综合征中观察到的异常相似。因此,hfh-4的靶向突变提供了一个模型,用于阐明调节纤毛发育和左右不对称确定的机制。