Identification and expression analysis of alternative splice variants of the rat Hax-1 gene
Identification and expression analysis of alternative splice variants of the rat Hax-1 gene
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DOI：
10.1016/j.gene.2005.11.035
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发表时间：
2006-04-12
期刊：
影响因子：
3.5
通讯作者：
Siedlecki, JA
中科院分区：
文献类型：
--
作者：
Grzybowska, EA;Sarnowska, E;Siedlecki, JA
Hax-1 蛋白已在小鼠和人类身上进行了研究，显示出有效的抗凋亡活性，并参与细胞运动的调节。大鼠 Hax-1 cDNA 的克隆揭示了 7 个替代转录物，其主要差异在于 5' 区域。替代剪接涉及外显子 1（在 5 个转录物中被跳过）、内含子 1（部分保留在这些转录物中）、外显子 2（可以部分跳过）和内含子 2（在一个变体中保留）。通过外显子连接特异性 RT-PCR 和 RNase 保护测定证实了不同剪接变体的存在。表达分析表明，总体 Hax-1 mRNA 水平在大多数组织中相对较低，而在睾丸中非常高，并且变体在不同组织中的表达模式相似。不同转录本的存在意味着存在多种蛋白质亚型，具有三个假定的起始密码子。通过蛋白质印迹证实了至少三种蛋白质亚型的存在。有趣的是，睾丸中的高 mRNA 水平并不转化为高蛋白质水平，这表明存在组织特异性翻译调节或受调节的蛋白质降解。 (c) 2006 Elsevier B.V. 保留所有权利。
Hax-1 protein, which has been studied in mice and humans, shows a potent anti-apoptotic activity and is involved in regulation of cell motility. Cloning of the rat Hax-1 cDNA has revealed seven alternative transcripts, which differ mostly in their 5' region Alternative splicing concerns exon 1, skipped in 5 transcripts, intron I which is partially retained in these transcripts, exon 2, which can be partially skipped, and intron 2, retained in one variant. The existence of different splicing variants was confirmed by exon-junction-specific RT-PCR and RNase protection assay. Analysis of expression indicates that overall Hax-1 mRNA level is relatively low in most tissues and very high in testes, and that the expression pattern of the variants is similar in different tissues. Presence of different transcripts implies the existence of several protein isoforms, with three putative start codons. The existence of at least three protein isoforms was confirmed by Western blot. Interestingly, high mRNA level in testes does not translate into high protein level, suggesting the existence of tissue-specific translational regulation or regulated protein degradation. (c) 2006 Elsevier B.V. All rights reserved.