Alterations of the transcriptome of Sulfolobus acidocaldarius by exoribonuclease aCPSF2.
Alterations of the transcriptome of Sulfolobus acidocaldarius by exoribonuclease aCPSF2.
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DOI：
10.1371/journal.pone.0076569
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Bläsi U
中科院分区：
文献类型：
--
作者：
Märtens B;Amman F;Manoharadas S;Zeichen L;Orell A;Albers SV;Hofacker I;Bläsi U
最近的研究在crenarchaeon Sulfolobus solfataricus（Sso）中鉴定了一种5 '到3'的核糖核酸外切酶，称为Sso-RNase J，已被重新分类为aCPSF 2（古细菌裂解和多聚腺苷酸化特异性因子2）组的β-CASP蛋白。在这项研究中，Sso-aCPSF 2的酸热硫化叶菌（Saci-aCPSF 2）的直系同源物的功能进行了表征。与Sso-aCPSF 2一样，Saci-aCPSF 2在体外以5“至3”方向性降解RNA。为了解决Saci-aCPSF 2的生物学意义，构建了缺失突变体，并采用高通量RNA测序评估了Saci-aCPSF 2对转录组谱的影响。该分析揭示了560个具有差异转录丰度的基因，表明该酶在RNA代谢中具有相当大的作用。此外，生物信息学分析揭示了几种优先在5 '端降解的转录本。这通过Northern印迹分析对两种转录本进行了示例性验证，首次表明aCPSF 2蛋白在古细菌泉古菌分支中的5'至3'方向mRNA衰变中发挥作用。
Recent studies identified a 5´ to 3´ exoribonuclease termed Sso-RNase J in the crenarchaeon Sulfolobus solfataricus (Sso), which has been reclassified to the aCPSF2 (archaeal cleavage and polyadenylation specificity factor 2) group of β-CASP proteins. In this study, the Sso-aCPSF2 orthologue of Sulfolobus acidocaldarius (Saci-aCPSF2) was functionally characterized. Like Sso-aCPSF2, Saci-aCPSF2 degrades RNA with 5´ to 3´ directionality in vitro. To address the biological significance of Saci-aCPSF2, a deletion mutant was constructed, and the influence of Saci-aCPSF2 on the transcriptome profile was assessed employing high throughput RNA sequencing. This analysis revealed 560 genes with differential transcript abundance, suggesting a considerable role of this enzyme in RNA metabolism. In addition, bioinformatic analyses revealed several transcripts that are preferentially degraded at the 5´ end. This was exemplarily verified for two transcripts by Northern-blot analyses, showing for the first time that aCPSF2 proteins play a role in 5' to 3' directional mRNA decay in the crenarchaeal clade of Archaea.
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