Analysis of the gene that encodes the complement regulatory protein, membrane inhibitor of reactive lysis (CD59). Identification of an alternatively spliced exon and characterization of the transcriptional regulatory regions of the promoter.
Analysis of the gene that encodes the complement regulatory protein, membrane inhibitor of reactive lysis (CD59). Identification of an alternatively spliced exon and characterization of the transcriptional regulatory regions of the promoter.
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DOI：
10.4049/jimmunol.157.4.1659
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发表时间：
1996-08
影响因子：
4.4
通讯作者：
M. Holguin;Carol B. Martin;T. Eggett;Charles J. Parker
中科院分区：
文献类型：
--
作者：
M. Holguin;Carol B. Martin;T. Eggett;Charles J. Parker
反应性溶解膜抑制因子（MIRL，CD 59）是一种18 kDa的糖基磷脂酰肌醇锚定蛋白，调节补体的膜攻击复合物的形成。这些研究的目的是表征编码CD 59的基因。我们的实验重新定义了基因的结构组织，通过识别以前未被识别的选择性剪接外显子。来自各种培养的人细胞系和PBMC的cDNA的PCR分析表明，含有可变剪接外显子序列的转录本与缺乏该序列的转录本一致表达。引物延伸研究表明，选择性剪接的CD 59 mRNA的转录起始位点与没有选择性剪接的外显子序列的转录物的转录起始位点相同，这表明两种形式的CD 59 mRNA的表达受到类似的调节。启动子区的分析表明，CD 59基因转录起始位点5'端的前70个核苷酸对于组成型转录和PMA响应性转录都是必需的;然而，对PMA的响应性是细胞系特异性的。总之，这些研究重新定义了CD 59基因的组织，并确定了参与组成型和PMA诱导型转录的启动子区域。
Membrane inhibitor of reactive lysis (MIRL, CD59) is an 18-kDa glycosylphosphatidylinositol-anchored protein that regulates formation of the membrane attack complex of complement. The purpose of these studies was to characterize the gene that encodes CD59. Our experiments redefined the structural organization of the gene by identifying a previously unrecognized alternatively spliced exon. Analysis by PCR of cDNA derived from a variety of cultured human cell lines and from PBMC showed that transcripts containing the alternatively spliced exon sequence were expressed concordantly with transcripts lacking that sequence. Primer extension studies demonstrated that the transcriptional start site of alternatively spliced CD59 mRNA is the same as that of transcripts without the alternatively spliced exon sequence, suggesting that expression of both forms of CD59 mRNA is regulated similarly. Analysis of the promoter region showed that the first 70 nucleotides immediately 5' of the transcriptional start site of the CD59 gene are essential for both constitutive and PMA-responsive transcription; however, responsiveness to PMA is cell line specific. Together, these studies have redefined the organization of the CD59 gene and identified regions of the promoter involved in constitutive and PMA-inducible transcription.