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Ccr4-Not去腺苷酸化酶复合体构成了秀丽隐杆线虫中主要的聚腺苷酸去除活性。

The Ccr4-Not deadenylase complex constitutes the main poly(A) removal activity in C. elegans.

作者信息

Nousch Marco, Techritz Nora, Hampel Daniel, Millonigg Sophia, Eckmann Christian R

机构信息

Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Pfotenhauerstrasse 108, 01307 Dresden, Germany.

出版信息

J Cell Sci. 2013 Sep 15;126(Pt 18):4274-85. doi: 10.1242/jcs.132936. Epub 2013 Jul 10.

Abstract

Post-transcriptional regulatory mechanisms are widely used to control gene expression programs of tissue development and physiology. Controlled 3' poly(A) tail-length changes of mRNAs provide a mechanistic basis of such regulation, affecting mRNA stability and translational competence. Deadenylases are a conserved class of enzymes that facilitate poly(A) tail removal, and their biochemical activities have been mainly studied in the context of single-cell systems. Little is known about the different deadenylases and their biological role in multicellular organisms. In this study, we identify and characterize all known deadenylases of Caenorhabditis elegans, and identify the germ line as tissue that depends strongly on deadenylase activity. Most deadenylases are required for hermaphrodite fertility, albeit to different degrees. Whereas ccr-4 and ccf-1 deadenylases promote germline function under physiological conditions, panl-2 and parn-1 deadenylases are only required under heat-stress conditions. We also show that the Ccr4-Not core complex in nematodes is composed of the two catalytic subunits CCR-4 and CCF-1 and the structural subunit NTL-1, which we find to regulate the stability of CCF-1. Using bulk poly(A) tail measurements with nucleotide resolution, we detect strong deadenylation defects of mRNAs at the global level only in the absence of ccr-4, ccf-1 and ntl-1, but not of panl-2, parn-1 and parn-2. Taken together, this study suggests that the Ccr4-Not complex is the main deadenylase complex in C. elegans germ cells. On the basis of this and as a result of evidence in flies, we propose that the conserved Ccr4-Not complex is an essential component in post-transcriptional regulatory networks promoting animal reproduction.

摘要

转录后调控机制被广泛用于控制组织发育和生理过程中的基因表达程序。mRNA的3' 多聚腺苷酸(poly(A))尾长度的可控变化为此类调控提供了一个机制基础,影响着mRNA的稳定性和翻译能力。去腺苷酸化酶是一类保守的酶,可促进多聚腺苷酸尾的去除,其生化活性主要在单细胞系统中进行研究。对于不同的去腺苷酸化酶及其在多细胞生物中的生物学作用,人们了解甚少。在本研究中,我们鉴定并表征了秀丽隐杆线虫所有已知的去腺苷酸化酶,并确定生殖系是强烈依赖去腺苷酸化酶活性的组织。大多数去腺苷酸化酶对雌雄同体的生育能力是必需的,尽管程度不同。在生理条件下,ccr-4和ccf-1去腺苷酸化酶促进生殖系功能,而panl-2和parn-1去腺苷酸化酶仅在热应激条件下是必需的。我们还表明,线虫中的Ccr4-Not核心复合物由两个催化亚基CCR-4和CCF-1以及结构亚基NTL-1组成,我们发现NTL-1可调节CCF-1的稳定性。使用具有核苷酸分辨率的大量多聚腺苷酸尾测量方法,我们仅在没有ccr-4、ccf-1和ntl-1的情况下,而非在没有panl-2、parn-1和parn-2的情况下,在全局水平检测到mRNA的强烈去腺苷酸化缺陷。综上所述,本研究表明Ccr4-Not复合物是秀丽隐杆线虫生殖细胞中的主要去腺苷酸化酶复合物。基于此以及果蝇中的证据,我们提出保守的Ccr4-Not复合物是促进动物繁殖的转录后调控网络中的一个重要组成部分。

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