The Role of a New Cleavage and Polyadenylation Specificity Factor (CPSF73-II)

新的切割和聚腺苷酸化特异性因子 (CPSF73-II) 的作用

基本信息

批准号：
7992573
负责人：
QINGSHUN Q LI
金额：
$ 5.81万
依托单位：
MIAMI UNIVERSITY OXFORD
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-28 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7992573
关键词：
3&apos Untranslated Regions Address Affect Animals Arabidopsis Arts Biochemical Biochemical Genetics Biological Models Cells Cellular biology Complex DNA Microarray Chip Data Development Embryonic Development Eukaryota Female Gene Expression Gene Expression Regulation Genes Germ Cells Goals Health Histones Human Link Maps Messenger RNA Microarray Analysis Molecular Biology Molecular Genetics Mutation Analysis Nature Organism Play Poly(A) Tail Polyadenylation Process Proteins RNA Polymerase II RNA Processing Reaction Research Research Personnel Role Specificity Spermatogenesis Students System Testing Transcript Translations Universities Untranslated Regions Virus Diseases Work base disorder control domain mapping endonuclease gene function genome sequencing interest mRNA Cleavage and Polyadenylation Factors mRNA Precursor novel tumor

项目摘要

DESCRIPTION (provided by applicant): The 3' untranslated region (UTR) and the poly(A) tail of an mRNA can exert regulatory influences that affect message stability, rate of translation, and/or subcellular localization. Thus, the choice of where the cleavage reaction takes place in a pre-mRNA can have significant influence on its subsequent activity and fate. Abnormalities in the process of 3'-end formation have been linked to viral infection, tumor development, and spermatogenesis in human and animals. The activity of the Cleavage and Polyadenylation Specificity Factor (CPSF) complex is required for the cleavage and polyadenylation of most RNA polymerase II-synthesized pre-mRNA transcripts. Analysis of genome sequences from multicellular organisms shows that there are paralogues of some of the genes whose protein products comprise CPSF, but the role(s) these gene products play in mRNA 3'-end formation is very much unknown. The goal of this proposal is to understand how one of these paralogues (CPSF73-II) works to regulate 3'- end formation of mRNAs via a combination of biochemical and molecular genetic means using Arabidopsis as the model system. CPSF73-II encodes a protein with a domain similar to an endonuclease implicated in the cleavage of pre-mRNA and some histone mRNAs. Mutation analysis of the Arabidopsis atCPSF73-II gene demonstrates an essential role in female gamete and embryo development. Our hypothesis, based on extensive preliminary data, is that this group of proteins are involved in developmentally regulated mRNA 3'-end formation which is distinguishable from the constitutive one. To test this hypothesis, we propose two specific aims. The results of the research will uncover how this essential component of mRNA 3'-end formation regulates gene expression. Aim 1: To map the interacting domains of two atCPSF73 proteins with atCPSF100 subunit to elucidate the nature of the complex. Aim 2: To reveal target mRNAs which are subject to CPSF73-II activity using microarray technology. to Health Genes encoding a novel protein CPSF73-II that may play an essential role in the development are being studied. This protein is highly conserved among multicellular organisms. Built on the understanding of the gene function in the model system of Arabidopsis, the first goal of the project is to understand the proteins interaction partners, the results of which will reveal how it works in the cell. Using the state-of-the- art DNA microarray approach, the second goal is to uncover the targets that are regulated by the protein. The results of this research will be applicable to other higher eukaryotes, including humans, in understanding the gene's contributions in messenger RNA processing which is a critical step in gene expression and regulation. The latter is known to be relevant to many human health and disease control issues.

描述（由申请人提供）：3'未翻译区域（UTR）和mRNA的poly（a）尾巴会发挥调节影响，影响信息稳定性，翻译速率和/或亚细胞定位。因此，在前MRNA中进行切割反应发生的位置可能会对其随后的活性和命运产生重大影响。在3'末期形成过程中的异常与人和动物的病毒感染，肿瘤发育和精子发生有关。裂解和聚腺苷酸化特异性因子（CPSF）复合物的活性是大多数RNA聚合酶II合成前MRNA转录本的裂解和聚腺苷酸化所必需的。对多细胞生物的基因组序列的分析表明，某些基因的近代人的蛋白质产物包含CPSF，但是这些基因产物在mRNA 3'-end形成中起的作用是鲜为人知的。该提案的目的是了解这些旁系同源物之一（CPSF73-II）如何通过使用拟南芥作为模型系统的生化和分子遗传手段的结合来调节mRNA的3'-末端形成。 CPSF73-II编码具有类似于与前MRNA和某些组蛋白mRNA裂解的内切酶类似的域的蛋白质。拟南芥ATCPSF73-II基因的突变分析在女性配子和胚胎发育中表现出至关重要的作用。我们基于广泛的初步数据的假设是，这组蛋白质参与了发育调节的mRNA 3'末端形成，这与本构的分歧是可以区分的。为了检验这一假设，我们提出了两个具体目标。该研究的结果将发现mRNA 3'-end形成的基本成分如何调节基因表达。 AIM 1：绘制与ATCPSF100亚基的两个ATCPSF73蛋白的相互作用结构域以阐明复合物的性质。目标2：揭示使用微阵列技术受到CPSF73-II活性的目标mRNA。对于编码一种新型蛋白质CPSF73-II的健康基因可能在开发中起着至关重要的作用。该蛋白在多细胞生物中是高度保守的。基于对拟南芥模型系统中基因功能的理解，该项目的第一个目标是了解蛋白质相互作用伙伴，其结果将揭示其在细胞中的工作方式。使用最先进的DNA微阵列方法，第二个目标是揭示受蛋白质调节的靶标。这项研究的结果将适用于包括人类在内的其他较高的真核生物，以理解基因在Messenger RNA加工中的贡献，这是基因表达和调节的关键步骤。后者已知与许多人类健康和疾病控制问题有关。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Arabidopsis mRNA polyadenylation machinery: comprehensive analysis of protein-protein interactions and gene expression profiling.

DOI：
10.1186/1471-2164-9-220
发表时间：
2008-05-14
期刊：
BMC genomics
影响因子：
4.4
作者：
Hunt AG;Xu R;Addepalli B;Rao S;Forbes KP;Meeks LR;Xing D;Mo M;Zhao H;Bandyopadhyay A;Dampanaboina L;Marion A;Von Lanken C;Li QQ
通讯作者：
Li QQ

Arabidopsis PCFS4, a homologue of yeast polyadenylation factor Pcf11p, regulates FCA alternative processing and promotes flowering time.