人类I型干扰素受体基因的转录终止/多聚腺苷化发生在多个位点。

L L Hensley, P J Vojta, V K Han, D C Lee, H Ozer
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引用次数: 1

摘要

基于先前报道的序列,我们分离了一个独立的cDNA克隆,编码人类I型干扰素受体(IFN-R)的结合成分。该cDNA与已发表的序列完全相同,除了缺少5'未翻译序列的62个碱基,并且终止于两个潜在聚腺苷化位点中的第一个位点。在对ifn敏感和ifn耐药的Daudi细胞的poly(A)+ rna的Northern blot分析中,该克隆cDNA杂交的mRNA主要为2.4 kb, mRNA长度为1.8、4.8和5.6 kb,偶尔也有6.9 kb。这些不同的转录本,在Raji B细胞和两个t细胞系Jurkat和MOLT-4中也观察到相似的水平,在高强度洗涤后,通过与cDNA亚片段对应的交替探针检测到。相比之下,只有4.8 kb和5.6 kb的转录本杂交到聚合酶链反应(PCR)衍生的探针上,该探针与第二个多聚腺苷化位点下游的基因组序列相对应。这些结果表明,由于在已知的任何一个聚腺苷化位点上的转录不完全终止,后一种转录本与主要的2.4 kb mRNA来自同一基因。最后,对总rna的Northern blot分析显示,在人类妊娠中期胎儿的许多组织中存在显性的2.4 kb I型IFN-R转录本,这表明I型IFN-R基因在多种细胞类型中组成性表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Transcription termination/polyadenylation occurs at multiple sites in the human type I interferon receptor gene.

Based on the previously reported sequence, we isolated an independent cDNA clone encoding a binding component of the human type I interferon receptor (IFN-R). This cDNA is identical to the published sequence except that it lacks 62 bases of 5' untranslated sequence and terminates at the first of two potential polyadenylation sites. In Northern blot analyses of poly(A)+RNAs from both IFN-sensitive and IFN-resistant Daudi cells, this cloned cDNA hybridized to a predominant mRNA of 2.4 kb, as well as to mRNAs of 1.8, 4.8, and 5.6 kb, and occasionally 6.9 kb. These various transcripts, which were also observed at similar levels in Raji B cells and two T-cell lines, Jurkat and MOLT-4, were detected after high-stringency washes, and by alternate probes corresponding to subfragments of the cDNA. In contrast, only the 4.8- and 5.6-kb transcripts hybridized to a polymerase chain reaction (PCR)-derived probe that corresponded to genomic sequences immediately down-stream from the second polyadenylation site. These results indicate that the latter transcripts arise from the same gene as the predominant 2.4-kb mRNA due to incomplete transcription termination at either of the known polyadenylation sites. Finally, Northern blot analysis of total RNAs revealed the presence of the predominant 2.4-kb type I IFN-R transcript in numerous tissues from second trimester human fetuses, suggesting that the type I IFN-R gene is constitutively expressed in multiple cell types.

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