Molecular cloning and expression of alternatively spliced PITSLRE protein kinase isoforms.
Molecular cloning and expression of alternatively spliced PITSLRE protein kinase isoforms.
复制标题
DOI：
10.1016/s0021-9258(17)40749-6
复制
发表时间：
1994-06
期刊：
影响因子：
--
通讯作者：
J. Xiang;J. Lahti;J. Grenet;J. Easton;V. Kidd
中科院分区：
文献类型：
--
作者：
J. Xiang;J. Lahti;J. Grenet;J. Easton;V. Kidd
p58 GTA蛋白激酶的最小异位表达导致涉及细胞周期延迟、有丝分裂灾难和细胞活力降低的挑衅性表型。此外，这种激酶在进化上是很保守的，广泛表达，其基因定位于人类1号染色体上的一个位置，在肿瘤发生的晚期经常缺失。在这里，我们报告的p58 GTA蛋白激酶是一个更大的蛋白质亚家族的成员。编码这些蛋白质的mRNA通过选择性剪接从多个重复基因产生。这些亚型的大小范围为50至110 kDa。选择性剪接亚型之间的差异定位于分子的氨基末端区域。整个p58 GTA开放阅读框架在大多数这些p58 GTA同种型中是保守的。预测序列的较大的亚型编码二分核定位信号序列和广泛的聚谷氨酸结构域。使用p58 GTA同种型的抗体来确认不同细胞类型中可变剪接同种型的存在，以及鉴定似乎由单独基因产生的两种另外的同种型。细胞分级分离研究表明，其中一种亚型仅在细胞核中发现，其余的在细胞质和细胞核中发现。一些p58 GTA亚型的表达和定位表明它们可能具有专门的细胞功能。由于从多个基因产生的大量同种型，我们建议根据p34 cdc 2激酶特有的PSTAIRE盒的保守序列和它们转录的基因将这些激酶命名为PITSLRE α 1、α 2-1、α 2-2、α 2-3、α 2-4、β 1、β 2-1和β 2-2。
Minimal ectopic expression of the p58GTA protein kinase results in a provocative phenotype involving cell cycle delay, mitotic catastrophe, and decreased cell viability. In addition, this kinase is well conserved evolutionarily, ubiquitously expressed, and its genes map to a position on human chromosome 1 frequently deleted in the late stages of tumorigenesis. Here we report that the p58GTA protein kinase is a member of a larger subfamily of proteins. The mRNAs encoding these proteins are generated by alternative splicing from multiple duplicated genes. These isoforms range in size from 50 to 110 kDa. Divergence between the alternatively spliced isoforms is localized to the amino-terminal region of the molecule. The entire p58GTA open reading frame is conserved in most of these p58GTA isoforms. The predicted sequences of the larger isoforms encode bipartite nuclear localization signal sequences and extensive polyglutamic acid domains. Antibodies to the p58GTA isoform were used to confirm the presence of the alternatively spliced isoforms in different cell types as well as identify two additional isoforms that appear to arise from a separate gene(s). Cellular fractionation studies indicate that one of the isoforms is found only in the nucleus, and the remainder are found in both the cytoplasm and the nucleus. Expression and localization of some p58GTA isoforms suggest that they may have specialized cellular functions. Because of the large number of isoforms generated from multiple genes we propose naming these kinases PITSLRE alpha 1, alpha 2-1, alpha 2-2, alpha 2-3 alpha 2-4, beta 1, beta 2-1, and beta 2-2 based on the conserved sequence of the PSTAIRE box unique to p34cdc2 kinases and the gene from which they are transcribed.