Regulation of mRNA processing: Mechanisms and Consequences

mRNA 加工的调控：机制和后果

基本信息

批准号：
10621295
负责人：
James L. Manley
金额：
$ 79.43万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-08 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621295
关键词：
Affect Amyotrophic Lateral Sclerosis Anemia Antiviral Response Area Brain Complex Coupled Cytoplasm Development Disease Event GATA1 gene Genes Genetic Transcription Goals Homeostasis Human Lead MAP3K7 gene Malignant Neoplasms Membrane Proteins Messenger RNA Motor Cortex Mutation NF-kappa B Neoplasms Neurodegenerative Disorders Nuclear Nuclear Inner Membrane Nuclear Pore Complex Nuclear Pore Complex Proteins Open Reading Frames Pathway interactions Patients Physiological Poly A Polyadenylation Pre-mRNA Polyadenylation Factor Process Protein Isoforms Proteins RNA RNA Helicase RNA Splicing Reaction Regulation SARS-CoV-2 infection Sampling Specificity Suggestion Translating Work cell type cofactor design exosome experimental study frontotemporal lobar dementia amyotrophic lateral sclerosis human embryonic stem cell insight mRNA Precursor p38 Mitogen Activated Protein Kinase permissiveness sporadic amyotrophic lateral sclerosis

项目摘要

Synthesis of eukaryotic mRNAs is a complex process, and includes splicing and 3’ end formation of mRNA precursors. My lab has studied these processes for many years, including recently how they function in differentiation and disease. This proposal continues our studies on these topics, and can be divided into the following areas. mRNA processing in cancer. Work investigating how mutations in genes encoding splicing factors lead to MDS and other neoplasms will be continued, with an emphasis on SF3B1. Studies to elucidate both the mechanism(s) by which SF3B1 mutations affect splicing as well as the pathways that are dysregulated and lead to disease will be pursued. With respect to mechanism, an immediate goal will be to continue characterizing the SF3B1-interacting region of SUGP1. Another goal is to identify the SUGP1- interacting RNA helicase and determine its function in normal BP recognition, which will then allow determination of the detailed mechanism by which SF3B1 mutations disrupt splicing. With respect to pathways, experiments to elucidate the details of the recently described MAP3K7/p38/GATA1 pathway that underlies anemia in MDS will be performed. Missplicing events that affect other relevant pathways, such as aberrant activation of NF-kB, will be studied. FUS and other RBPs in ALS/FTD. With respect to FUS, experiments to analyze FUS nucleocytoplasmic homeostasis, a process important for formation of toxic cytoplasmic FUS aggregates, will be continued. Evidence supporting a gating mechanism involving the NPC and interactions with specific nucleoporins, themselves implicated in amyotrophic lateral sclerosis (ALS), will be pursued. Suggestions that this process may involve cell-type specificity, with MNs perhaps being more permissive, will be investigated. Experiments pursing the observation that RBP aggregates and consequent missplicing occur in a large fraction of sporadic ALS/FTD patient brains in the absence of known mutations will be continued. Aggregates will be isolated from motor cortex samples and protein/RNA composition determined to investigate what might nucleate their formation. Regulation of PA factor activity by AS. Experiments investigating the functions of isoforms of the polyadenylation (PA) factor WDR33 will be pursued. Two short isoforms, v2 and v3, are produced by intronic PA), and v2 but not v3 is an inner nuclear membrane protein not directly involved in PA. Interacting proteins of both will be identified, and results suggesting they are upregulated by an NF-kB pathway will be pursued. Evidence that the two isoforms function in the antiviral response, including to SARS-CoV-2 infection, will be further investigated. The finding that 45 isoforms of the PA factor Fip1 are produced in humans will be explored. eRNAs as mRNAs. Results suggesting that certain nuclear unstable lncRNAs, including eRNAs, are stabilized, exported to the cytoplasm and in some case translated, will be pursued. Observations that up to 5% of eRNAs contain significant ORFs and may be translated when the exosome cofactor Mtr4 is depleted will be further investigated. The possible physiological significance of eRNA “activation” will be studied in differentiated hESCs, which were shown to have sharply reduced levels of Mtr4.

真核mRNA的合成是一个复杂的过程，包括剪接和3'mRNA的末端形成前体。我的实验室已经研究了这些过程多年了，包括最近它们在分化和疾病。该建议继续我们对这些主题的研究，可以分为以下区域。癌症中的mRNA处理。研究研究如何编码剪接的基因中的突变导致MD和其他肿瘤的因素将继续，重点是SF3B1。阐明的研究 SF3B1突变影响剪接的机制以及途径将追求失调并导致疾病。关于机制，直接目标是继续表征SUGP1的SF3B1相互作用区域。另一个目标是确定SUGP1- 相互作用的RNA解旋酶并确定其在正常BP识别中的功能，然后将允许 SF3B1突变破坏剪接的详细机制的确定。关于途径阐明最近描述的MAP3K7/p38/GATA1途径的实验将执行MDS中的贫血。错过影响其他相关途径的事件，例如异常 NF-KB的激活将研究。 ALS/FTD中的FUS和其他RBP。关于FUS，实验分析FUS核细胞质稳态，这对于形成有毒细胞质FUS的过程很重要聚合将继续。支持门控机制的证据涉及NPC和相互作用对于特定的核蛋白，将追求在肌萎缩性侧索硬化症（ALS）中实施。建议这个过程可能将研究涉及细胞类型的特异性（MN）可能会更允许。实验追求RBP聚集物和随之而来的失误发生的观察结果很大一部分在没有已知突变的情况下，ALS/FTD患者大脑将继续。聚集体将与运动皮层样品和蛋白质/RNA成分确定要研究可能核能是什么形成。通过AS调节PA因子活性。研究了研究的功能将追求多腺苷酸化（PA）因子WDR33。两个简短的同工型V2和V3由内含子产生 PA）和V2而非V3是一种内部核膜蛋白，与PA无直接参与。相互作用的蛋白质两者都将被识别，结果表明，将追求NF-KB途径进行更新。抗病毒反应中两种同工型功能（包括SARS-COV-2感染）的证据将是进一步调查。在人类中产生了45个PA因子Fip1的同工型的发现将是探索。 Ernas作为mRNA。结果表明，包括Ernas在内的某些核不稳定的LNCRNA是稳定，出口到细胞质，在某些情况下翻译，将被追捕。观察到高达5％ ERNAS包含重要的ORF，当外泌体辅因子Mtr4耗尽时可能会翻译进一步调查。 ERNA“激活”可能的身体意义将在分化的hESC被证明急剧降低了MTR4的水平。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The E592K variant of SF3B1 creates unique RNA missplicing and associates with high-risk MDS without ring sideroblasts.

DOI：
10.21203/rs.3.rs-2802265/v1
发表时间：
2023-04-14
期刊：
Research square
影响因子：
0
作者：
Choi, In Young;Ling, Jonathan;Dalton, W Brian
通讯作者：
Dalton, W Brian

Nuclear RNA transcript levels modulate nucleocytoplasmic distribution of ALS/FTD-associated protein FUS.

DOI：
10.1038/s41598-022-12098-4
发表时间：
2022-05-17
期刊：
Scientific reports
影响因子：
4.6
作者：
通讯作者：

The RNA polymerase II CTD "orphan" residues: Emerging insights into the functions of Tyr-1, Thr-4, and Ser-7.

RNA 聚合酶 II CTD“孤儿”残基：对 Tyr-1、Thr-4 和 Ser-7 功能的新见解。

DOI：
10.1080/21541264.2017.1338176
发表时间：
2018
期刊：
Transcription
影响因子：
0
作者：
Yurko,NathanM;Manley,JamesL
通讯作者：
Manley,JamesL

Replication protein A associates with nucleolar R loops and regulates rRNA transcription and nucleolar morphology.

DOI：
10.1101/gad.348858.121
发表时间：
2021-12-01
期刊：
Genes & development
影响因子：
10.5
作者：
Feng S;Manley JL
通讯作者：
Manley JL

C9orf72 and triplet repeat disorder RNAs: G-quadruplex formation, binding to PRC2 and implications for disease mechanisms.

C9orf72 和三联体重复紊乱 RNA：G 四链体形成、与 PRC2 的结合以及对疾病机制的影响。

DOI：
10.1261/rna.071191.119
发表时间：
2019
期刊：
RNA (New York, N.Y.)
影响因子：
0
作者：
Wang,Xueyin;Goodrich,KarenJ;Conlon,ErinG;Gao,Jianchao;Erbse,AnnetteH;Manley,JamesL;Cech,ThomasR
通讯作者：
Cech,ThomasR