Bachmann M, Hilker M, Grölz D, Tellmann G, Hake U, Kater L, de Wilde P, Tröster H
Institut für Physiologische Chemie, Johannes-Gutenberg Universität, Mainz, Germany.
J Autoimmun. 1996 Dec;9(6):757-66. doi: 10.1006/jaut.1996.0098.
Recently we isolated a La/SS-B mRNA isoform from a cDNA library made from peripheral blood lymphocytes of a patient with primary Sjögren's Syndrome. In the La/SS-B mRNA isoform the exon 1 was replaced. The alternative exon was termed exon 1'. Genomic analysis showed that the exon 1' La mRNA was the result of a promoter-switch in combination with alternative splicing. Due to the unusual structure of the exon 1' La/SS-B mRNA, the function and the behaviour under physiological and pathophysiological conditions in tissue of patients with primary Sjögren's syndrome or Systemic Lupus Erythematosus remained obscure. Therefore assays were established allowing a qualitative and quantitative estimation of expression of the exon 1 and 1' La mRNA form, including in situ and dot blot hybridization as well as reversed PCR. Both mRNA forms were found to represent finally processed cytoplasmic mRNAs belonging to the abundant class of mRNAs. They were expressed and regulated in parallel. A ratio exon 1 to 1' between 1:1 and 5:1 was determined. Both mRNA forms were downregulated in quiescent cells and upregulated in activated and proliferating cells including non-keratized stratified squamous epithelial, endothelial, salivary gland as well as infiltrating cells.
最近,我们从一名原发性干燥综合征患者外周血淋巴细胞构建的cDNA文库中分离出一种La/SS - B mRNA异构体。在该La/SS - B mRNA异构体中,外显子1被替换。这个替代外显子被命名为外显子1'。基因组分析表明,外显子1'的La mRNA是启动子转换与可变剪接共同作用的结果。由于外显子1'的La/SS - B mRNA结构异常,其在原发性干燥综合征或系统性红斑狼疮患者组织中的生理和病理生理条件下的功能及行为仍不清楚。因此,我们建立了一些检测方法,可对包括原位杂交、斑点杂交以及逆转录PCR在内的外显子1和1'的La mRNA形式的表达进行定性和定量评估。发现这两种mRNA形式最终均代表属于丰富mRNA类别的加工成熟的细胞质mRNA。它们的表达和调控是平行的。确定外显子1与1'的比例在1:1至5:1之间。在静止细胞中这两种mRNA形式均下调,而在包括非角化复层鳞状上皮细胞、内皮细胞、唾液腺细胞以及浸润细胞在内的活化和增殖细胞中上调。