Ribosome profiling of the retrovirus murine leukemia virus.
Ribosome profiling of the retrovirus murine leukemia virus.
复制标题
逆转录病毒鼠白血病病毒的核糖体分析。
DOI：
10.1186/s12977-018-0394-5
复制
发表时间：
2018-01-22
期刊：
影响因子：
3.3
通讯作者：
Firth AE
中科院分区：
文献类型：
--
作者：
Irigoyen N;Dinan AM;Brierley I;Firth AE
逆转录病毒鼠白血病病毒（MuLV）具有8.3kb的RNA基因组，具有简单的5′-gag-pol-env-3′结构。pol基因的翻译依赖于gag UAG终止密码子的通读;而env基因从剪接的mRNA转录物翻译。在这里，我们报告了通过MuLV感染细胞的串联核糖体分析（RiboSeq）和RNA测序（RNASeq）对逆转录病毒基因表达的首次高分辨率分析。使用翻译抑制剂三尖杉酯碱和放线菌酮对MuLV感染的细胞进行核糖体分析，以分别区分起始和延长核糖体。宿主细胞基因表达的荟萃分析表明，RiboSeq数据集以高分辨率特异性捕获了翻译核糖体的足迹。MuLV基因组RNA的核糖体占有率的直接测量表明，约7%的核糖体经历gag终止密码子通读以进入pol基因。发现翻译起始发生在gag和env转录本5′前导序列内的几个额外位点，位于它们各自注释的起始密码子的上游。这些实验揭示了MuLV基因组内存在许多先前未表征的、核糖体占据的开放阅读框，可能具有调控后果。此外，我们提供了细胞感染期间终止密码子通读效率的第一个直接测量。本文的在线版本（10.1186/s12977-018-0394-5）包含补充材料，可供授权用户使用。
The retrovirus murine leukemia virus (MuLV) has an 8.3 kb RNA genome with a simple 5′-gag-pol-env-3′ architecture. Translation of the pol gene is dependent upon readthrough of the gag UAG stop codon; whereas the env gene is translated from spliced mRNA transcripts. Here, we report the first high resolution analysis of retrovirus gene expression through tandem ribosome profiling (RiboSeq) and RNA sequencing (RNASeq) of MuLV-infected cells. Ribosome profiling of MuLV-infected cells was performed, using the translational inhibitors harringtonine and cycloheximide to distinguish initiating and elongating ribosomes, respectively. Meta-analyses of host cell gene expression demonstrated that the RiboSeq datasets specifically captured the footprints of translating ribosomes at high resolution. Direct measurement of ribosomal occupancy of the MuLV genomic RNA indicated that ~ 7% of ribosomes undergo gag stop codon readthrough to access the pol gene. Initiation of translation was found to occur at several additional sites within the 5′ leaders of the gag and env transcripts, upstream of their respective annotated start codons. These experiments reveal the existence of a number of previously uncharacterised, ribosomally occupied open reading frames within the MuLV genome, with possible regulatory consequences. In addition, we provide the first direct measurements of stop codon readthrough efficiency during cellular infection. The online version of this article (10.1186/s12977-018-0394-5) contains supplementary material, which is available to authorized users.
登录
查看更多内容
通过RNA测序和核糖体分析对冠状病毒基因表达的高分辨率分析。
影响因子：
6.7
作者：
Irigoyen N;Firth AE;Jones JD;Chung BY;Siddell SG;Brierley I
通讯作者：
Brierley I
影响因子：
6.4
作者：
Ahi YS;Zhang S;Thappeta Y;Denman A;Feizpour A;Gummuluru S;Reinhard B;Muriaux D;Fivash MJ;Rein A
通讯作者：
Rein A
DOI：
10.1261/rna.052548.115
发表时间：
2015-10
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Chung BY;Hardcastle TJ;Jones JD;Irigoyen N;Firth AE;Baulcombe DC;Brierley I
通讯作者：
Brierley I
影响因子：
14.9
作者：
Nakagawa S;Niimura Y;Gojobori T;Tanaka H;Miura K
通讯作者：
Miura K
影响因子：
5.4
作者：
Hibbert, CS;Mirro, J;Rein, A
通讯作者：
Rein, A