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规范多聚(A)聚合酶 PAP1 对非编码 RNA 进行多聚腺苷酸化,是布氏锥虫 snoRNA 生物发生所必需的。

The Canonical Poly (A) Polymerase PAP1 Polyadenylates Non-Coding RNAs and Is Essential for snoRNA Biogenesis in Trypanosoma brucei.

机构信息

The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar-Ilan University, Ramat-Gan 5290002, Israel.

Department of Epidemiology and Microbial Diseases, Yale School of Public Health, New Haven, CT 06536, USA.

出版信息

J Mol Biol. 2017 Oct 27;429(21):3301-3318. doi: 10.1016/j.jmb.2017.04.015. Epub 2017 Apr 26.

Abstract

The parasite Trypanosoma brucei is the causative agent of African sleeping sickness and is known for its unique RNA processing mechanisms that are common to all the kinetoplastidea including Leishmania and Trypanosoma cruzi. Trypanosomes possess two canonical RNA poly (A) polymerases (PAPs) termed PAP1 and PAP2. PAP1 is encoded by one of the only two genes harboring cis-spliced introns in this organism, and its function is currently unknown. In trypanosomes, all mRNAs, and non-coding RNAs such as small nucleolar RNAs (snoRNAs) and long non-coding RNAs (lncRNAs), undergo trans-splicing and polyadenylation. Here, we show that the function of PAP1, which is located in the nucleus, is to polyadenylate non-coding RNAs, which undergo trans-splicing and polyadenylation. Major substrates of PAP1 are the snoRNAs and lncRNAs. Under the silencing of either PAP1 or PAP2, the level of snoRNAs is reduced. The dual polyadenylation of snoRNA intermediates is carried out by both PAP2 and PAP1 and requires the factors essential for the polyadenylation of mRNAs. The dual polyadenylation of the precursor snoRNAs by PAPs may function to recruit the machinery essential for snoRNA processing.

摘要

寄生虫布氏锥虫是非洲昏睡病的病原体,以其独特的 RNA 处理机制而闻名,这种机制在所有动基体目生物中都很常见,包括利什曼原虫和克氏锥虫。锥虫拥有两种典型的 RNA 多聚(A)聚合酶(PAPs),分别称为 PAP1 和 PAP2。PAP1 由该生物中仅有的两个含有顺式剪接内含子的基因之一编码,其功能目前尚不清楚。在锥虫中,所有的 mRNA 和非编码 RNA,如小核仁 RNA(snoRNA)和长非编码 RNA(lncRNA),都经历反式剪接和多聚腺苷酸化。在这里,我们表明位于核内的 PAP1 的功能是多聚腺苷酸化经历反式剪接和多聚腺苷酸化的非编码 RNA。PAP1 的主要底物是 snoRNA 和 lncRNA。在 PAP1 或 PAP2 沉默的情况下,snoRNA 的水平降低。snoRNA 中间体的双重多聚腺苷酸化由 PAP2 和 PAP1 共同完成,并需要 mRNA 多聚腺苷酸化所必需的因子。PAPs 对前体 snoRNA 的双重多聚腺苷酸化可能有助于招募 snoRNA 加工所必需的机制。

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