Emetine blocks DNA replication via proteosynthesis inhibition not by targeting Okazaki fragments.
Emetine blocks DNA replication via proteosynthesis inhibition not by targeting Okazaki fragments.
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依然氨酸通过蛋白质合成抑制而不是通过靶向冈崎片段来阻止DNA复制。
DOI：
10.26508/lsa.202201560
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发表时间：
2022-09-09
影响因子：
4.4
通讯作者：
Moudry, Pavel
中科院分区：
文献类型：
--
作者：
Lukac, David;Machacova, Zuzana;Moudry, Pavel
这项研究澄清了长期以来埃米丁作为滞后链合成特异性抑制剂的滥用。依美汀通过抑制蛋白质生物合成来阻断两条DNA链的合成。领先和滞后链的DNA合成是独立工作的，如果受到RPA分子的保护，细胞可以耐受在解链过程中产生的单链DNA。天然生物碱埃美汀被用作滞后链合成、解偶联引导和滞后链复制的特异性抑制剂。在这里，通过对滞后链合成抑制剂的分析，我们表明，尽管依美汀完全抑制DNA复制：它不会诱导单链DNA和染色质结合RPA32(CB-RPA32)的产生。与此一致的是，依美汀不会激活复制检查点，也不会激活DNA损伤反应。依美汀也是蛋白质合成的抑制剂，持续的蛋白质合成对于DNA的准确复制是必不可少的。从机制上讲，我们证明埃美汀对蛋白质合成的急性阻断在时间上先于其对DNA复制的影响。因此，我们的结果与艾美汀通过抑制蛋白质合成影响DNA复制的假设是一致的。依美汀和轻微的POLA1抑制可阻止S相聚(腺苷二磷酸核糖)。总而言之，我们的研究表明埃米丁不是一种特定的滞后链合成抑制剂，这意味着它在分子生物学中的使用。
This study clarifies the long-standing misuse of emetine as a lagging strand synthesis-specific inhibitor. Emetine blocks the synthesis of both DNA strands by the inhibition of protein biosynthesis. DNA synthesis of the leading and lagging strands works independently and cells tolerate single-stranded DNA generated during strand uncoupling if it is protected by RPA molecules. Natural alkaloid emetine is used as a specific inhibitor of lagging strand synthesis, uncoupling leading and lagging strand replication. Here, by analysis of lagging strand synthesis inhibitors, we show that despite emetine completely inhibiting DNA replication: it does not induce the generation of single-stranded DNA and chromatin-bound RPA32 (CB-RPA32). In line with this, emetine does not activate the replication checkpoint nor DNA damage response. Emetine is also an inhibitor of proteosynthesis and ongoing proteosynthesis is essential for the accurate replication of DNA. Mechanistically, we demonstrate that the acute block of proteosynthesis by emetine temporally precedes its effects on DNA replication. Thus, our results are consistent with the hypothesis that emetine affects DNA replication by proteosynthesis inhibition. Emetine and mild POLA1 inhibition prevent S-phase poly(ADP-ribosyl)ation. Collectively, our study reveals that emetine is not a specific lagging strand synthesis inhibitor with implications for its use in molecular biology.
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DOI：
10.1083/jcb.200806068
发表时间：
2008-12-29
期刊：
The Journal of cell biology
影响因子：
--
作者：
Sugimura K;Takebayashi S;Taguchi H;Takeda S;Okumura K
通讯作者：
Okumura K
影响因子：
64.5
作者：
HANDELI, S;KLAR, A;CEDAR, H
通讯作者：
CEDAR, H
影响因子：
11.2
作者：
Xiao, Gu;Lundine, Devon;Bargonetti, Jill
通讯作者：
Bargonetti, Jill
影响因子：
16.6
作者：
Thakar, Tanay;Leung, Wendy;Moldovan, George-Lucian
通讯作者：
Moldovan, George-Lucian
DOI：
10.1038/nrm.2017.53
发表时间：
2017-10
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Ray Chaudhuri A;Nussenzweig A
通讯作者：
Nussenzweig A