MOLECULAR-CLONING OF A DEVELOPMENTALLY-REGULATED BRAIN PROTEIN, CHICKEN DREBRIN-A AND ITS EXPRESSION BY ALTERNATIVE SPLICING OF THE DREBRIN GENE
MOLECULAR-CLONING OF A DEVELOPMENTALLY-REGULATED BRAIN PROTEIN, CHICKEN DREBRIN-A AND ITS EXPRESSION BY ALTERNATIVE SPLICING OF THE DREBRIN GENE
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DOI：
10.1016/0169-328x(93)90154-h
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发表时间：
1993-07-01
期刊：
影响因子：
--
通讯作者：
OBATA, K
中科院分区：
文献类型：
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作者：
KOJIMA, N;SHIRAO, T;OBATA, K
Drebrins是在鸡脑中发现的发育调节蛋白，分为E1、E2和A三种形式。以前我们从鸡胚cDNA文库中分离到两个与胚胎发育相关的cDNA。它们的不同之处在于，一个内部129个核苷酸的序列，命名为ins1，被插入到编码dreplastin E2的cDNA中，而在另一个编码dreplastin E1的cDNA中被删除。为了寻找编码dreplasma A的cDNA克隆，以胚胎dreplasma cDNA片段为探针，从1日龄鸡脑cDNA文库中筛选出dreplasma A基因。因此，分离出一种新的cDNA，其序列与dreplastin E2的序列完全相同，只是在5 ′方向上插入了一个138个核苷酸的序列，称为ins2，它紧接着ins1的上游。由于整个编码区的翻译产物与dreplastin A的翻译产物相似，因此该cDNA应对应于dreplastin A的mRNA。对3个drebrins基因cDNA的序列分析表明，鸡drebrins基因的异质性是由ins1和ins2两个序列的插入或缺失引起的。氨基端半区包括ins2和两个短序列在羧基端的预测dreplastin A是高度进化保守的。dreplastin基因的克隆和测序显示，ins1和ins2独立编码的单独的外显子和三个dreplastin异构体被认为是由选择性剪接从一个单一的dreplastin基因。在表达的时间过程和组织分布的每个dreplastin的差异表明，dreplastin基因的选择性剪接位点选择的机制是在一个发育阶段依赖性和组织特异性的方式进行调节。
Drebrins are developmentally regulated proteins found in the chicken brain and are classified into three forms, E1, E2 and A. Previously we isolated two cDNAs corresponding to the embryonic drebrin mRNAs from a chick embryo cDNA library. They differed in that an internal 129-nucleotide sequence, designated ins1, was inserted in the cDNA encoding drebrin E2 and was deleted in the other cDNA encoding drebrin E1. To search for the cDNA clone encoding drebrin A, a cDNA library of 1-day-old chick brains was screened using embryonic drebrin cDNA fragments as probes. Consequently, a novel cDNA was isolated, the sequence of which was entirely identical with that of drebrin E2 except for the insertion of a 138-nucleotide sequence, designated ins2, in the 5' direction immediately upstream from ins1. Since the translation product of the entire coding region was similar to that of drebrin A, this cDNA should correspond to the mRNA for drebrin A. Sequencing analysis of three drebrin cDNAs clearly indicated that the heterogeneity of chicken drebrins was caused by the insertion or deletion of the two sequences, ins1 and ins2. The amino-terminal half region including ins2 and two short sequences in the carboxyl-terminal region of the predicted drebrin A were highly evolutionarily conserved. Cloning and sequencing of the drebrin gene revealed that ins1 and ins2 were independently encoded by separate exons and three drebrin isoforms were thought to arise by alternative splicing from a single drebrin gene. The difference in the time course of expression and tissue distribution of each drebrin suggests that the machinery of alternative splicing site selection of the drebrin gene is regulated in a developmental stage-dependent and tissue-specific manner.