Human eosinophil Charcot-Leyden crystal protein: cloning and characterization of a lysophospholipase gene promoter.
Human eosinophil Charcot-Leyden crystal protein: cloning and characterization of a lysophospholipase gene promoter.
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DOI：
10.1182/blood.v82.6.1868.1868
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发表时间：
1993
期刊：
影响因子：
20.3
通讯作者：
H. Gomolin;Y. Yamaguchi;A. V. Paulpillai;L. Dvorak;S. Ackerman;D. Tenen
中科院分区：
文献类型：
--
作者：
H. Gomolin;Y. Yamaguchi;A. V. Paulpillai;L. Dvorak;S. Ackerman;D. Tenen
Charcot-Leyden晶体（CLC）蛋白是一种仅由嗜酸性粒细胞和嗜碱性粒细胞表达的溶血磷脂酶。嗜酸性细胞系在嗜酸性细胞分化过程中，CLC稳态mRNA水平显著升高。这种增加的表达在丁酸盐诱导早幼粒细胞白血病HL-60细胞系嗜酸性亚群（C15）的过程中受到转录调节，正如核运行试验所示。CLC基因的转录起始位点位于最长cDNA序列5′端上游43 bp处。从19号染色体特异性文库中克隆了编码CLC蛋白的基因，并对其转录起始位点重叠的片段进行了分离和测序。在HL-60-C15细胞以及其他髓系（U937）和非髓系（HeLa和RPMI 8402）细胞系的瞬时转染中，质粒构建（在pXP2荧光素酶表达载体中）在CLC转录起始位点上游含有411和292 bp基因组序列，从而指导报告基因的表达。然而，两种CLC启动子结构在这些细胞系中的差异表达表明，启动子的-292至- 411bp区域可能在嗜酸性粒细胞谱系中具有一定的表达特异性。CLC启动子序列包含两个一致的GATA结合位点，一个富含嘌呤的序列是ets基因家族成员PU.1的潜在结合位点，以及其他髓细胞特异性启动子中描述的序列。这是功能性嗜酸性粒细胞启动子的首次证明，该启动子可以作为鉴定嗜酸性粒细胞谱系在承诺和分化过程中调节基因表达的DNA元件和反式激活因子的模型。
The Charcot-Leyden crystal (CLC) protein is a lysophospholipase expressed exclusively by eosinophils and basophils. During eosinophilic differentiation of eosinophil-committed cell lines, CLC steady state mRNA levels increase significantly. This increased expression is transcriptionally regulated during butyrate induction of an eosinophilic subline (C15) of the promyelocytic leukemia cell line HL-60, as shown by nuclear run-on assays. The transcriptional start site of the CLC gene was identified 43 bp upstream of the 5' end of the longest available cDNA sequence. The gene encoding CLC protein was cloned from a chromosome 19-specific library and a fragment overlapping the transcriptional start site was isolated and sequenced. Plasmid constructs (in the pXP2 luciferase expression vector) containing 411 and 292 bp of genomic sequence upstream of the CLC transcriptional start site directed reporter gene expression in transient transfections of HL-60-C15 cells, as well as other myeloid (U937) and nonmyeloid (HeLa and RPMI 8402) cell lines. However, the differential expression of the two CLC promoter constructs in these cell lines suggests that the -292 to -411 bp region of the promoter may confer some specificity for expression in the eosinophil lineage. The CLC promoter sequence contains two consensus GATA binding sites, a purine-rich sequence that presents potential binding sites for PU.1, a member of the ets family of genes, as well as sequences described in other myeloid-specific promoters. This is the first demonstration of a functional eosinophil promoter that could serve as a model for identifying DNA elements and trans-activating factors that regulate gene expression during the commitment and differentiation of the eosinophil lineage.