Participation of Proteins of the CPSF Complex in Polyadenylation of Transcripts Read by RNA Polymerase III from SINEs

IF 1.5 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biology Pub Date : 2024-06-05 DOI:10.1134/s0026893324700122

I. G. Ustyantsev, O. R. Borodulina, D. A. Kramerov

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Abstract

SINEs are mobile genetic elements of multicellular eukaryotes that arose during evolution from various tRNAs, as well as from 5S rRNA and 7SL RNA. Like the genes of these RNAs, SINEs are transcribed by RNA polymerase III. The transcripts of some mammalian SINEs have the capability of AAUAAA-dependent polyadenylation, which is unique for transcript generated by RNA polymerase III. Despite a certain similarity with canonical polyadenylation of mRNAs (transcripts of RNA polymerase II), these processes apparently differ significantly. The purpose of this work is to evaluate how important for polyadenylation of SINE transcripts are proteins of the CPSF complex formed by mPSF and mCF subcomplexes which direct mRNA polyadenylation. In HeLa cells, siRNA knockdowns of the CPSF components were carried out, after which the cells were transfected with plasmid constructs containing SINEs. A decrease in polyadenylation of the SINE transcripts as a result of the knockdown of the proteins was evaluated by Northern-hybridization. It turned out that the CPSF components, such as Wdr33 and CPSF30, contributed to the polyadenylation of SINE transcriptions, while the knockdown of CPSF100, CPSF73, and symplekin did not reduce the polyadenylation of these transcripts. Wdr33 and CPSF30, along with the CPSF160 and Fip1 previously studied, are components of the subcomplex mPSF responsible for mRNA polyadenylation. Thus, the available data suggest the importance of all mPSF proteins for polyadenylation of SINE transcripts. At the same time, CPSF100, CPSF73, and symplekin, forming the subcomplex mCF, are responsible for the cleavage of pre-mRNA; therefore, their non-participation in the polyadenylation of SINE transcriptions seems quite natural.

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CPSF 复合物中的蛋白质参与了由 RNA 聚合酶 III 从 SINEs 读取的转录本的多腺苷酸化过程

摘要SINEs是多细胞真核生物的移动遗传元件，在进化过程中从各种tRNA以及5S rRNA和7SL RNA中产生。与这些 RNA 的基因一样，SINEs 也是由 RNA 聚合酶 III 转录的。一些哺乳动物 SINE 的转录本具有 AAUAAA 依赖性多腺苷酸化功能，这是由 RNA 聚合酶 III 生成的转录本所独有的。尽管与 mRNA（RNA 聚合酶 II 的转录本）的典型多腺苷酸化有一定的相似性，但这些过程显然有很大的不同。这项工作的目的是评估由 mPSF 和 mCF 亚复合物形成的 CPSF 复合物中的蛋白质对 SINE 转录本的多聚腺苷酸化有多重要，这些蛋白质可指导 mRNA 的多聚腺苷酸化。在 HeLa 细胞中，用 siRNA 敲除 CPSF 成分，然后用含有 SINEs 的质粒构建体转染细胞。通过 Northern-hybridization 方法评估了 SINE 转录本的多腺苷酸化程度是否因蛋白质被敲除而降低。结果发现，Wdr33和CPSF30等CPSF成分有助于SINE转录本的多腺苷化，而CPSF100、CPSF73和sympplekin的敲除并没有减少这些转录本的多腺苷化。Wdr33 和 CPSF30 以及之前研究的 CPSF160 和 Fip1 是负责 mRNA 多腺苷化的亚复合体 mPSF 的组成成分。因此，现有数据表明，所有 mPSF 蛋白对 SINE 转录本的多腺苷酸化都很重要。同时，组成亚复合体 mCF 的 CPSF100、CPSF73 和 symplekin 负责前 mRNA 的裂解；因此，它们不参与 SINE 转录本的多腺苷酸化似乎很自然。

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来源期刊

Molecular Biology 生物-生化与分子生物学

CiteScore

1.30

自引率

8.30%

发文量

审稿时长

3 months

期刊介绍： Molecular Biology is an international peer reviewed journal that covers a wide scope of problems in molecular, cell and computational biology including genomics, proteomics, bioinformatics, molecular virology and immunology, molecular development biology, molecular evolution and related areals. Molecular Biology publishes reviews, experimental and theoretical works. Every year, the journal publishes special issues devoted to most rapidly developing branches of physical-chemical biology and to the most outstanding scientists.