Small RNA analysis in Sindbis virus infected human HEK293 cells.
Small RNA analysis in Sindbis virus infected human HEK293 cells.
复制标题
DOI：
10.1371/journal.pone.0084070
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Powell PP
中科院分区：
文献类型：
--
作者：
Donaszi-Ivanov A;Mohorianu I;Dalmay T;Powell PP
与植物和无脊椎动物中RNA干扰（RNAi）的防御机制相反，它在哺乳动物对病毒感染的先天反应中的作用是一个有争议的问题。由于RNAi在控制甲病毒属辛德毕斯病毒（SINV）在昆虫中的感染中具有公认的作用，因此我们使用该病毒来研究RNAi在SINV感染人类细胞中的作用。使SINV AR 339和TR 339-GFP适应于在HEK 293细胞中生长。SINV感染早期（4和6 hpi）的小RNA（sRNA）深度测序显示病毒sRNA（vsRNA）丰度低（0.8%），没有Dicer产物的大小、序列或位置特异性模式，也没有任何可识别的模式可归因于特定的RNAi生物发生途径。这得到了每个序列的多个变体以及沿着病毒基因组序列沿着缺乏热点的支持。最佳定义的vsRNA的丰度低于北方印迹检测的限度。病毒在HEK 293细胞中的适应性实验表明，与对照相比，病毒的序列变化不大，但E1基因存在一个从G到C的SNP位点。通过测序显示出一些微小差异表达的12种miRNA，通过北方印迹分析显示表达没有差异。我们发现，与无脊椎动物的SINV感染不同，Dicer依赖性svRNA的产生和细胞miRNA表达的变化并没有被检测到作为人类对SINV反应的一部分。
In contrast to the defence mechanism of RNA interference (RNAi) in plants and invertebrates, its role in the innate response to virus infection of mammals is a matter of debate. Since RNAi has a well-established role in controlling infection of the alphavirus Sindbis virus (SINV) in insects, we have used this virus to investigate the role of RNAi in SINV infection of human cells. SINV AR339 and TR339-GFP were adapted to grow in HEK293 cells. Deep sequencing of small RNAs (sRNAs) early in SINV infection (4 and 6 hpi) showed low abundance (0.8%) of viral sRNAs (vsRNAs), with no size, sequence or location specific patterns characteristic of Dicer products nor did they possess any discernible pattern to ascribe to a specific RNAi biogenesis pathway. This was supported by multiple variants for each sequence, and lack of hot spots along the viral genome sequence. The abundance of the best defined vsRNAs was below the limit of Northern blot detection. The adaptation of the virus to HEK293 cells showed little sequence changes compared to the reference; however, a SNP in E1 gene with a preference from G to C was found. Deep sequencing results showed little variation of expression of cellular microRNAs (miRNAs) at 4 and 6 hpi compared to uninfected cells. Twelve miRNAs exhibiting some minor differential expression by sequencing, showed no difference in expression by Northern blot analysis. We show that, unlike SINV infection of invertebrates, generation of Dicer-dependent svRNAs and change in expression of cellular miRNAs were not detected as part of the Human response to SINV.
登录
查看更多内容
影响因子：
13.5
作者：
Hou, Weihong;Tian, Qing;Zheng, Jianyu;Bonkovsky, Herbert L.
通讯作者：
Bonkovsky, Herbert L.
影响因子：
3.7
作者：
Burke, Crystal W.;Gardner, Christina L.;Steffan, Joshua J.;Ryman, Kate D.;Klimstra, William B.
通讯作者：
Klimstra, William B.
影响因子：
6.7
作者：
Kim Y;Lee S;Kim S;Kim D;Ahn JH;Ahn K
通讯作者：
Ahn K
影响因子：
4.2
作者：
Campbell CL;Keene KM;Brackney DE;Olson KE;Blair CD;Wilusz J;Foy BD
通讯作者：
Foy BD
Mirbase：MicroRNA基因组学的工具。
影响因子：
14.9
作者：
Griffiths-Jones, Sam;Saini, Harpreet Kaur;van Dongen, Stijn;Enright, Anton J.
通讯作者：
Enright, Anton J.