Hippe Andreas, Bylaite Matilda, Chen Min, von Mikecz Anna, Wolf Ronald, Ruzicka Thomas, Walz Markus
Department of Dermatology, Heinrich-Heine University, Moorenstrasse 5, D-40225 Duesseldorf, Germany.
Gene. 2006 Sep 1;379:116-26. doi: 10.1016/j.gene.2006.04.027. Epub 2006 May 13.
HAX1 is an ubiquitously expressed human gene. Though a number of cellular and viral proteins are known to interact with HAX1, its function is still not completely understood. On the basis of these identified interaction partners, HAX1 seems to play a role in apoptosis and the organization of the cytoskeleton. The cDNAs for human and mouse Hax1 share 86% identity and 80% identity at the protein level, suggesting a similar functional importance. To date, no conclusive data on the tissue specific expression of the murine Hax1 are available and only one interaction partner has been identified. Here, we show a detailed expression analysis for the murine ortholog by RT-PCR, Northern and Western blot. Furthermore, the distribution of Hax1 within different mouse tissues was studied by immunohistochemistry (IHC). In general, we found a good correlation between the results obtained from different detection techniques. Similar to its human counterpart, mouse Hax1 seems to be ubiquitously expressed. At the RNA level, we found the highest expression of Hax1 in liver, kidney and testis. In sharp contrast to the human HAX1 which is highly expressed in skeletal muscle, the mouse ortholog was detected only at very low levels. Using a specific antibody, we detected Hax1 in the majority of mouse tissues by IHC. Interestingly, the most prominent expression of Hax1 was found in epithelial, endothelial and muscle cells. Surprisingly, thymus, spleen and pancreas did not show detectable immunostaining. Furthermore, we have studied the subcellular localisation of Hax1 in a keratinocyte and a neuronal cell line by immunofluorescence. We found Hax1 to be localised mainly in the cytoplasm and detected a partial colocalisation with mitochondria. The results presented here summarize for the first time the expression of the murine Hax1 in different tissues and two cell lines. Further studies will elucidate the functional importance of this protein in individual cell types with respect to structural aspects, cell mobility and apoptosis.
HAX1是一种在人体中普遍表达的基因。尽管已知许多细胞和病毒蛋白可与HAX1相互作用,但其功能仍未完全明确。基于这些已确定的相互作用伙伴,HAX1似乎在细胞凋亡和细胞骨架组织中发挥作用。人类和小鼠Hax1的cDNA在核酸水平上有86%的同源性,在蛋白质水平上有80%的同源性,这表明它们在功能上具有相似的重要性。迄今为止,关于小鼠Hax1的组织特异性表达尚无确凿数据,仅鉴定出一个相互作用伙伴。在此,我们通过逆转录聚合酶链反应(RT-PCR)、Northern印迹法和Western印迹法对小鼠直系同源基因进行了详细的表达分析。此外,我们还通过免疫组织化学(IHC)研究了Hax1在不同小鼠组织中的分布。总体而言,我们发现不同检测技术所得结果之间具有良好的相关性。与人类对应物相似,小鼠Hax1似乎也在全身广泛表达。在RNA水平上,我们发现Hax1在肝脏、肾脏和睾丸中表达最高。与在骨骼肌中高表达的人类HAX1形成鲜明对比的是,小鼠直系同源基因的表达水平极低。我们使用一种特异性抗体,通过免疫组织化学在大多数小鼠组织中检测到了Hax1。有趣的是,Hax1最显著的表达出现在上皮细胞、内皮细胞和肌肉细胞中。令人惊讶的是,胸腺、脾脏和胰腺未显示出可检测到的免疫染色。此外,我们通过免疫荧光研究了Hax1在角质形成细胞和神经元细胞系中的亚细胞定位。我们发现Hax1主要定位于细胞质中,并检测到其与线粒体有部分共定位。本文呈现的结果首次总结了小鼠Hax1在不同组织和两种细胞系中的表达情况。进一步的研究将阐明该蛋白在个体细胞类型中对于结构方面、细胞迁移和细胞凋亡的功能重要性。