Dissecting the in vivo assembly of the 30S ribosomal subunit reveals the role of RimM and general features of the assembly process.
Dissecting the in vivo assembly of the 30S ribosomal subunit reveals the role of RimM and general features of the assembly process.
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剖析 30S 核糖体亚基的体内组装揭示了 RimM 的作用和组装过程的一般特征
DOI：
10.1093/nar/gks1256
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发表时间：
2013-02-01
影响因子：
14.9
通讯作者：
Gao N
中科院分区：
文献类型：
--
作者：
Guo Q;Goto S;Chen Y;Feng B;Xu Y;Muto A;Himeno H;Deng H;Lei J;Gao N
核糖体生物合成是一个严格调控的多步骤过程。核糖体亚基的组装是复杂的生物发生过程的中心步骤，涉及细菌体内近30种蛋白质因子。虽然组装过程已经在体外广泛研究了40多年，但对于组装因子的体内过程和特定作用的信息非常有限。这样的例子是核糖体成熟因子M（RimM），一种参与30 S亚基后期组装的因子。在这里，我们结合定量质谱和冷冻电子显微镜表征在体内30 S组装中间体分离的突变大肠杆菌菌株与基因的组装因子删除。我们的组成和结构数据表明，在这些中间体中，30 S亚基的3′-结构域的组装严重延迟，与成熟的30 S亚基相比，具有高度代表性的3′-结构域蛋白和较大的构象差异。进一步的分析表明RimM不仅具有促进一些3′-结构域蛋白组装的功能，而且还具有稳定rRNA三级结构的功能。更重要的是，这项研究揭示了体外和体内组装途径之间有趣的相似性和不同之处，表明它们在总体上是相似的，但有细微的差异。
Ribosome biogenesis is a tightly regulated, multi-stepped process. The assembly of ribosomal subunits is a central step of the complex biogenesis process, involving nearly 30 protein factors in vivo in bacteria. Although the assembly process has been extensively studied in vitro for over 40 years, very limited information is known for the in vivo process and specific roles of assembly factors. Such an example is ribosome maturation factor M (RimM), a factor involved in the late-stage assembly of the 30S subunit. Here, we combined quantitative mass spectrometry and cryo-electron microscopy to characterize the in vivo 30S assembly intermediates isolated from mutant Escherichia coli strains with genes for assembly factors deleted. Our compositional and structural data show that the assembly of the 3′-domain of the 30S subunit is severely delayed in these intermediates, featured with highly underrepresented 3′-domain proteins and large conformational difference compared with the mature 30S subunit. Further analysis indicates that RimM functions not only to promote the assembly of a few 3′-domain proteins but also to stabilize the rRNA tertiary structure. More importantly, this study reveals intriguing similarities and dissimilarities between the in vitro and the in vivo assembly pathways, suggesting that they are in general similar but with subtle differences.
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影响因子：
7.4
作者：
Bunner, Anne E.;Trauger, Sunia A.;Siuzdak, Gary;Williamson, James R.
通讯作者：
Williamson, James R.
影响因子：
5.6
作者：
Dutca, Laura M.;Culver, Gloria M.
通讯作者：
Culver, Gloria M.
DOI：
10.1073/pnas.0912007107
发表时间：
2010-03-23
影响因子：
11.1
作者：
Bunner, Anne E.;Beck, Andrea H.;Williamson, James R.
通讯作者：
Williamson, James R.
影响因子：
4.5
作者：
Bubunenko, Mikhail;Korepanov, Alexey;Culver, Gloria M.
通讯作者：
Culver, Gloria M.
影响因子：
5.6
作者：
Gabashvili, IS;Agrawal, RK;Frank, J
通讯作者：
Frank, J