Transcription starting from an alternative promoter leads to the expression of the human ABO histo-blood group antigen
Transcription starting from an alternative promoter leads to the expression of the human ABO histo-blood group antigen
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DOI：
10.1046/j.1537-2995.2003.00382.x
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发表时间：
2003-05-01
期刊：
影响因子：
2.9
通讯作者：
Yamamoto, F
中科院分区：
文献类型：
--
作者：
Hata, Y;Kominato, Y;Yamamoto, F
背景技术背景：利用离体培养AC 133-CD 34（+）细胞的cDNA末端5 ′-快速扩增技术，最近在先前确定的转录起始位点上游约0.7kb处鉴定了转录起始位点。在红系和上皮细胞系的细胞中均显示了来自选择性起始外显子1a的转录物。由于外显子1a的核粒序列不包含ATG密码子，我们检测了从外显子1a开始的转录是否导致功能性糖基转移酶的产生。研究设计和方法：使用各种A转移酶表达质粒进行人胃癌MKN 28细胞的稳定转染实验。在用A转移酶表达质粒转染的细胞上证明了大量的A抗原，所述A转移酶表达质粒含有来自外显子1a的完整cDNA或5 ′-截短的cDNA，导致产生N-缺失胞质尾区和部分跨膜结构域的截短蛋白。然而，可忽略不计的数量的A抗原的A转移酶的表达质粒转染细胞含有5 '-截短的cDNA，导致生产的N-截短的蛋白质没有胞质尾和跨膜domain.CONCLUSION：这项研究表明，功能性A转移酶可以通过从外显子1a转录产生。
BACKGROUND: Using the 5'-rapid amplification of cDNA ends technique with the ex vivo culture of AC133-CD34(+) cells, a transcription start site was recently identified approximately 0.7 kb upstream from the transcription start sites previously determined. The transcripts from the alternative starting exon 1a were demonstrated in the cells of both erythroid and epithelial lineages. Because the nucleoticle sequence of exon la does not contain an ATG codon, we examined whether transcription starting from exon la leads to production of a functional glycosyltransferase.STUDY DESIGN AND METHODS: Stable transfection experiments into the human gastric cancer MKN28 cells were performed using the various A transferase expression plasmids.RESULTS: Large amounts of A antigens were demonstrated on the cells transfected with the A transferase expression plasmid containing the entire cDNA from exon 1a or the 5'-truncated cDNA leading to the production of the N-truncated protein with deletion of the cytoplasmic tail and a portion of the transmembrane domain. However, negligible amounts of A antigens were observed on the cells transfected with the A transferase expression plasmids containing the 5'-truncated cDNA leading to the production of the N-truncated proteins without the cytoplasmic tail and the transmembrane domain.CONCLUSION: This study suggests that a functional A transferase could be produced by the transcription from exon 1 a.