Yeast pre-rRNA processing and modification occur cotranscriptionally.
Yeast pre-rRNA processing and modification occur cotranscriptionally.
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DOI：
10.1016/j.molcel.2010.02.024
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发表时间：
2010-03-26
期刊：
影响因子：
16
通讯作者：
Tollervey D
中科院分区：
文献类型：
--
作者：
Kos M;Tollervey D
为了更好地了解酵母核糖体的合成，我们开发了代谢标记培养物的快速采集和分析技术。对得到的动力学数据进行建模，可以将预测的寿命和加工模式与实验数据进行比较。这支持∼170rRNA的35S初级转录本在30℃下的转录时间(∼40 NT S−1)，其中高比例(∼70%)的新生转录本在产生18S rRNA前体的20S前体的早期加工位点被切割。这种新生的转录本切割水平显然与之前的报道相冲突，即酵母Pre-rRNA的修饰仅发生在已发布的转录本上。第二轮高分辨率动力学标记显示，20S前rRNA主要作为新生转录本经历甲基化，而25S/5.8S rRNA的27S前体在新生转录本上部分甲基化。结果表明，对前rRNA加工的定量分析可以产生重要的生物学见解。►快速标记是为了为►合成的建模提供动力学数据，该模型显示了高度共转录的Pre-rRNA裂解►，这表明新转录的►也会发生rRNA修饰。进一步的标记证实了新生的Pre-rRNA转录本的大量甲基化
To better understand yeast ribosome synthesis, we developed techniques for the rapid harvesting and analysis of metabolically labeled cultures. Modeling of the resulting kinetic data allowed predicted lifetimes and processing patterns to be compared with the experimental data. This supported a transcription time for the 35S primary transcripts of ∼170 s at 30°C (∼40 nt s−1), with a high fraction (∼70%) of nascent transcripts cleaved at the early processing sites that generate the 20S precursor to the 18S rRNA. This level of nascent transcript cleavage apparently conflicted with previous reports that modification of yeast pre-rRNA exclusively occurred on released transcripts. A second round of high-resolution kinetic labeling showed that 20S pre-rRNA predominately undergoes methylation as nascent transcripts, whereas the 27S precursor to the 25S/5.8S rRNAs was partially methylated on the nascent transcript. The results demonstrate that quantitative analyses of pre-rRNA processing can yield important biological insights. ► Fast labeling was developed to provide kinetic data for modeling of rRNA synthesis ► The model indicated a high degree of cotranscriptional pre-rRNA cleavage ► This suggested that rRNA modification would also occur on nascent transcripts ► Further labeling confirmed substantial methylation of nascent pre-rRNA transcripts
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影响因子：
3.2
作者：
LACROUTE, F
通讯作者：
LACROUTE, F
影响因子：
16
作者：
Grandi, P;Rybin, V;Hurt, E
通讯作者：
Hurt, E
影响因子：
5.6
作者：
UDEM, SA;WARNER, JR
通讯作者：
WARNER, JR
DOI：
10.1111/j.1432-1033.1977.tb11531.x
发表时间：
1977-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BRAND, RC;KLOOTWIJK, J;PLANTA, RJ
通讯作者：
PLANTA, RJ
影响因子：
11.4
作者：
TOLLERVEY, D
通讯作者：
TOLLERVEY, D