Cloning of focal adhesion kinase, pp125(FAK), from rat brain reveals multiple transcripts with different patterns of expression
Cloning of focal adhesion kinase, pp125(FAK), from rat brain reveals multiple transcripts with different patterns of expression
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DOI：
10.1016/0169-328x(95)00273-u
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发表时间：
1996-04-01
期刊：
影响因子：
--
通讯作者：
Girault, JA
中科院分区：
文献类型：
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作者：
Burgaya, F;Girault, JA
粘着斑激酶（Focal adhesion kinase，pp 125，FAK）是一种富含粘着斑的细胞质酪氨酸激酶。我们筛选了一个大鼠纹状体cDNA文库与特异性FAK信使的PCR扩增的cDNA探针。多个克隆的测序揭示了由5个保守盒的组合产生的三个不同的5 '-前导序列的存在。其中一个包含一个潜在的替代起始位点，78个碱基对上游的先前描述。另一个与位于3号染色体上的人类基因组序列有89%相同。大多数阳性克隆在负责粘着斑靶向的区域中含有编码三个氨基酸（Pro-Trp-Arg）的插入。我们建议将这种蛋白质的变体命名为FAK +。用特异性引物和探针进行RT-PCR和Southern杂交，研究FAK多种形式的表达模式。不同的5 '-前导序列总是以相同的比例被发现。相比之下，FAK + mRNA在非神经组织中的表达水平非常低，而在所有脑区中均高度表达。在培养的细胞中，它存在于星形胶质细胞中，并在神经元中富集。这些结果表明存在多种形式的FAK转录本和蛋白，其中之一，FAK +，可能在神经系统中发挥特定的作用。
Focal adhesion kinase (pp125(FAK), or FAK) is a cytoplasmic tyrosine kinase enriched in focal adhesions. We have screened a rat striatum cDNA library with a PCR-amplified cDNA probe specific for FAK messenger. Sequencing of multiple clones revealed the existence of three different 5'-leader sequences resulting from the combination of 5 conserved boxes. One of them contains a potential alternative initiation site, 78 base pairs upstream of that previously described. Another is 89% identical to a human genomic sequence located on chromosome 3. Most positive clones contained an insertion coding for three amino acids (Pro-Trp-Arg) in the region responsible for focal adhesion targeting. We propose to name this variant of the protein FAK +. The pattern of expression of the multiple forms of FAK was studied by RT-PCR and Southern hybridization with specific primers and probes. The different 5'-leader sequences were always found in the same proportions. In contrast, FAK + mRNA was found at very low levels in non-nervous tissues, whereas it was highly expressed in all brain regions. In cells in culture, it was present in astrocytes and enriched in neurons. These results demonstrate the existence of multiple forms of FAK transcript and protein, one of which, FAK +, may play a specific role in the nervous system.