Dehlin E, von Gabain A, Alm G, Dingelmaier R, Resnekov O
Institute of Microbiology and Genetics, Vienna Biocenter, Austria.
Mol Cell Biol. 1996 Feb;16(2):468-74. doi: 10.1128/MCB.16.2.468.
Expression of beta interferon (IFN-beta) is transiently induced when Namalwa B cells (Burkitt lymphoma cell line) are infected by Sendai virus. In this study, we found that an elongation of the IFN-beta mRNA could be detected in virus-infected cells and that such a modification was not observed when the IFN-beta transcript was induced by a nonviral agent, poly(I-C). Treatment of the cells with a transcriptional inhibitor (actinomycin D or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole) resulted in further elongation of the transcript. Characterization of the elongated IFN-beta transcript by primer extension and RNase H treatment showed that the modification was a result of an elongated poly(A) tail of up to 400 nucleotides. We conclude that the poly(A) tail elongation of the IFN-beta transcript is associated with the viral infection. Furthermore, the presence of the elongated IFN-beta transcript correlated with a decrease of IFN-beta protein in the medium and in cell extracts. Sucrose gradient analysis of cytoplasmic extracts showed that IFN-beta transcripts with elongated poly(A) tails were found in the nonpolysomal fractions, whereas the shorter transcripts could be detected in both polysomal and nonpolysomal fractions. A longer form of the IFN-beta mRNA was also found in the nonpolysomal fractions of cells not treated with transcriptional inhibitors. Thus, the observed regulation of IFN-beta mRNA is not entirely dependent on the inhibition of transcription. To our knowledge, this study provides the first example of a poly(A) tail elongation in somatic cells that negatively influences gene expression in vivo.
当仙台病毒感染Namalwa B细胞(伯基特淋巴瘤细胞系)时,β干扰素(IFN-β)的表达会被短暂诱导。在本研究中,我们发现在病毒感染的细胞中可检测到IFN-β mRNA的延长,而当IFN-β转录本由非病毒剂聚肌苷酸-聚胞苷酸(poly(I-C))诱导时未观察到这种修饰。用转录抑制剂(放线菌素D或5,6-二氯-1-β-D-呋喃核糖基苯并咪唑)处理细胞导致转录本进一步延长。通过引物延伸和核糖核酸酶H处理对延长的IFN-β转录本进行表征表明,这种修饰是长达400个核苷酸的延长聚腺苷酸尾的结果。我们得出结论,IFN-β转录本的聚腺苷酸尾延长与病毒感染有关。此外,延长的IFN-β转录本的存在与培养基和细胞提取物中IFN-β蛋白的减少相关。细胞质提取物的蔗糖梯度分析表明,具有延长聚腺苷酸尾的IFN-β转录本存在于非多聚核糖体组分中,而较短的转录本可在多聚核糖体和非多聚核糖体组分中均检测到。在未用转录抑制剂处理的细胞的非多聚核糖体组分中也发现了较长形式的IFN-β mRNA。因此,观察到的IFN-β mRNA的调控并不完全依赖于转录抑制。据我们所知,本研究提供了体细胞中聚腺苷酸尾延长对体内基因表达产生负面影响的首个实例。