Kinetic analysis reveals the fate of a microRNA following target regulation in mammalian cells.
Kinetic analysis reveals the fate of a microRNA following target regulation in mammalian cells.
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DOI：
10.1016/j.cub.2011.01.067
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发表时间：
2011-03-08
期刊：
影响因子：
9.2
通讯作者：
Brown, Brian D.
中科院分区：
文献类型：
--
作者：
Baccarini, Alessia;Chauhan, Hemangini;Gardner, Thomas J.;Jayaprakash, Anitha D.;Sachidanandam, Ravi;Brown, Brian D.
关于microRNA(MiRNA)的生物发生和调控的相当多的细节已经被揭示，然而，对于靶调控之后miRNA的命运知之甚少。为了深入了解这一过程，我们对miRNA在生物发生终止后以及在调节不受Ago2介导的催化切割的靶的过程中的周转进行了动力学分析。通过对miRNA及其靶分子在稳态和衰变过程中分子数量的定量，我们发现每个miRNA分子至少能够调控2个靶转录本，这在体内提供了证据，证明miRNA与其靶分子不是不可逆转的隔离，并且miRNA调控的非切片途径是多次周转。通过深度测序，我们进一步证明了miRNA的循环受到靶调控的限制，靶调控促进了miRNA 3‘端的转录后修饰，并加快了miRNA的衰退速度。这些研究为miRNA调控的效率提供了新的见解，有助于解释miRNA如何调控大量转录本，并确定哺乳动物细胞中miRNA衰退的特异性机制之一。
Considerable details about microRNA (miRNA) biogenesis and regulation have been uncovered, however, little is known about the fate of the miRNA subsequent to target regulation. To gain insight into this process, we carried out kinetic analysis of a miRNA’s turnover following termination of its biogenesis, and during regulation of a target that is not subject to Ago2-mediated catalytic cleavage. By quantitating the number of molecules of the miRNA and its target in steady-state, and in the course of its decay, we found that each miRNA molecule was able to regulate at least 2 target transcripts, providing in vivo evidence that the miRNA is not irreversibly sequestered with its target, and that the non-slicing pathway of miRNA regulation is multiple-turnover. Using deep-sequencing, we further show that miRNA recycling is limited by target regulation, which promotes post-transcriptional modifications to the 3′ end of the miRNA, and accelerates the miRNA’s rate of decay. These studies provide new insight into the efficiency of miRNA regulation, which help to explain how a miRNA can regulate a vast number of transcripts, and identify one of the mechanisms that impart specificity to miRNA decay in mammalian cells.
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