Further characterisation of the translational termination-reinitiation signal of the influenza B virus segment 7 RNA.
Further characterisation of the translational termination-reinitiation signal of the influenza B virus segment 7 RNA.
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DOI：
10.1371/journal.pone.0016822
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发表时间：
2011-02-08
期刊：
影响因子：
3.7
通讯作者：
Brierley I
中科院分区：
文献类型：
--
作者：
Powell ML;Leigh KE;Pöyry TA;Jackson RJ;Brown TD;Brierley I
终止依赖性再起始用于协同调节双顺反子流感病毒B片段7 RNA的M1和BM 2开放阅读框（ORF）的表达。BM 2 ORF的起始密码子与M1 ORF的终止密码子在五核苷酸UAA UG中重叠，并且终止于M1终止密码子的约10%的核糖体在重叠的AUC处重新启动翻译。BM 2合成需要存在并翻译通过紧邻UAA UG上游的45 nt RNA，称为“终止上游核糖体结合位点”（TURBS）。该区域可以将核糖体40 S亚基在终止后拴系到mRNA，并且TURBS的短区域，基序1，与18 S rRNA的螺旋26互补，涉及该过程。在这里，我们提供了进一步的证据，mRNA和rRNA之间的直接相互作用，在酵母细胞中使用反义寡核苷酸靶向和功能分析。TURBS还结合起始因子eIF 3，我们在这里表明，这种蛋白质刺激重新启动从野生型和有缺陷的TURBS外源性时，可能通过稳定核糖体-mRNA的相互作用。此外，我们表明，TURBS相对于UAA UG重叠的位置是至关重要的，和终止太远下游的18 S互补序列抑制的过程中，可能是由于减少40 S拴系。然而，在报告基因mRNA中，只有重新启动密码子被移到下游，终止-重新启动被抑制但没有被消除，因此重新启动的位点有些灵活。在eIF 4G缺失的RRL中，远距离AUG上的再起始不受抑制，这表明拴系的40 S亚基可以移动一定距离而不需要线性扫描。
Termination-dependent reinitiation is used to co-ordinately regulate expression of the M1 and BM2 open-reading frames (ORFs) of the dicistronic influenza B segment 7 RNA. The start codon of the BM2 ORF overlaps the stop codon of the M1 ORF in the pentanucleotide UAA UG and ∼10% of ribosomes terminating at the M1 stop codon reinitiate translation at the overlapping AUG. BM2 synthesis requires the presence of, and translation through, 45 nt of RNA immediately upstream of the UAA UG, known as the ‘termination upstream ribosome binding site’ (TURBS). This region may tether ribosomal 40S subunits to the mRNA following termination and a short region of the TURBS, motif 1, with complementarity to helix 26 of 18S rRNA has been implicated in this process. Here, we provide further evidence for a direct interaction between mRNA and rRNA using antisense oligonucleotide targeting and functional analysis in yeast cells. The TURBS also binds initiation factor eIF3 and we show here that this protein stimulates reinitiation from both wild-type and defective TURBS when added exogenously, perhaps by stabilising ribosome-mRNA interactions. Further, we show that the position of the TURBS with respect to the UAA UG overlap is crucial, and that termination too far downstream of the 18S complementary sequence inhibits the process, probably due to reduced 40S tethering. However, in reporter mRNAs where the restart codon alone is moved downstream, termination-reinitiation is inhibited but not abolished, thus the site of reinitiation is somewhat flexible. Reinitiation on distant AUGs is not inhibited in eIF4G-depleted RRL, suggesting that the tethered 40S subunit can move some distance without a requirement for linear scanning.
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影响因子：
3.7
作者：
Napthine S;Lever RA;Powell ML;Jackson RJ;Brown TD;Brierley I
通讯作者：
Brierley I
影响因子：
5.4
作者：
McCormick, Christopher J.;Salim, Omar;Clarke, Ian N.
通讯作者：
Clarke, Ian N.
影响因子：
3.7
作者：
BRIEDIS, DJ;LAMB, RA;CHOPPIN, PW
通讯作者：
CHOPPIN, PW
影响因子：
11.4
作者：
Horvath, C M;Williams, M A;Lamb, R A
通讯作者：
Lamb, R A
影响因子：
5.4
作者：
Meyers, Gregor
通讯作者：
Meyers, Gregor