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Vitamin D and Estrogen Action in Bone: Concerted Role for hnRNPs in the C Family

维生素 D 和雌激素在骨中的作用：C 家族中 hnRNP 的协同作用

基本信息

批准号：
8514513
负责人：
John S Adams
金额：
$ 39.23万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-01-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8514513
关键词：
1,25 (OH) vitamin D Accident and Emergency department Adolescent Alternative Splicing Attention Beryllium Binding Binding Proteins Biological Assay Bone Diseases Bone Growth Cells Chromatin Coin DNA Binding Disease Outbreaks Dominant-Negative Mutation Drops Estradiol Estrogen Receptor 1 Estrogen Receptors Estrogens Event Family Female Adolescents Gene Expression Generations Genes Genetic Transcription Health Heterogeneous-Nuclear Ribonucleoprotein Group C Heterogeneous-Nuclear Ribonucleoproteins Hormone Receptor Hormones Human Humulus Immunoprecipitation Individual Investigation Left Ligands Link Los Angeles Messenger RNA Methodology MicroRNAs Modeling Mus Mutation Nature Osteoblasts Osteogenesis Participant Phenotype Precipitation Primates Process Production Proteins Public Health RNA RNA Binding RNA Splicing Research Resistance Response Elements Rickets Role Signal Pathway Single-Stranded DNA Site Skeleton Sterols Technology Testing Therapeutic Intervention Transcript Transgenic Organisms Translations Untranslated RNA Vehicle crash Vitamin D Vitamin D Response Element Vitamin D3 Receptor Work bone chromatin modification chromatin remodeling ds-DNA expectation hormone response element human female man novel nucleic acid binding protein overexpression receptor receptor binding research study small molecule steroid hormone theories young man

项目摘要

DESCRIPTION (provided by applicant): Twenty-five years ago we began to investigate an outbreak of rachitic bone disease in adolescent, female New World primates residing at the Los Angles Zoo. Our investigation of this "experiment of nature" and that of an adolescent human female with a similar phenotype led us to the discovery of a novel means for vitamin D and estrogen resistance in primates, including man. We coined these resistance-causing proteins the vitamin D response element binding protein (VDRE-BP) and estrogen response element binding (ERE-BP) for their ability to compete in trans with the liganded vitamin D receptor (VDR) and estrogen receptor 1 (ER1) for their cognate response elements, identifying them as nucleic acid binding proteins in the heterogeneous nuclear ribonucleoprotein C (hnRNPC) family. Recent work has led us to realize that the ability of these hnRNPs to alter steroid hormone-directed gene expression is not limited to their dominant-negative actions at the level of transcription. By virtue of their capacity to interact with single-strand DNA (ssDNA), ssRNA as well as double- strand DNA (dsDNA), these hnRNPs have the potential to exert control over gene expression at multiple sites in the cell. Here we theorize that a specific hnRNP can act as a multi-site participant in the synchronized expression of a single gene product by way of regulating, in succession, chromatin remodeling (ssDNA binding), transcription (dsDNA binding), splicing (ssDNA and ssRNA binding) and microRNA (miRNA; ssRNA binding). Because both the human and subhuman primate experiments of nature in hnRNP overexpression we have studied resulted in coincident vitamin D- and estrogen-antagonism on the growing skeleton of adolescent females, here we will concentrate on growing bone and the osteoblast as natural targets for these hnRNPs. Three specific aims are proposed: 1) using chromatin modification/precipitation, over/under expression and novel cell free translation technologies, explore the impact of hnRNPs on the cells responsible for 1,25- dihydroxyvitamin D and estradiol-driven bone growth; 2) employ new RNA immunoprecipitation and micro- RNA methodology to study the versatility of these hnRNPS to control transcript splicing and handling by directed binding to RNA products of the same gene whose expression is simultaneously regulated at the level of chromatin remodeling and transcription; and 3) ascertain the composite effect of these hnRNPs on the growing, adolescent skeleton by targeted transgenic overexpression of the VDRE-BP and ERE-BP hnRNPs in mouse bone. It is our expectation that the proposed research will pave the way to a mechanistic understanding of how these multi-functional hnRNPs control the process of sterol/steroid hormone-regulated gene expression in growing bone and delineate novel points of potential therapeutic intervention in that process. PUBLIC HEALTH RELEVANCE: Vitamin D and estrogen are two steroid hormones long recognized for their ability to influence the growing skeleton. Twenty five years ago we began to investigate an outbreak of rickets in adolescent, female New World primates residing at the Los Angeles Zoo, identifying proteins in the heterogeneous nuclear ribonucleoprotein C (hnRNPC) family, the vitamin D response element binding protein (VDRE-BP) and estrogen response element binding (ERE-BP) which caused resistance to the actions vitamin D and estrogen hormones on the skeleton. By virtue of their capacity to interact with single-strand DNA (ssDNA), ssRNA as well as double- strand DNA (dsDNA), we have planned experiments to test our theory that the VDRE-BP and ERE-BP can act in bone as a multi-site participant in the synchronized expression of a single gene product by way of regulating, in succession, i) chromatin remodeling, ii) transcription, iii) transcript splicing and iv) transcript handling.

描述(申请人提供)：25年前，我们开始调查在洛杉矶动物园居住的青少年女性新世界灵长类动物中爆发的脊椎骨病。我们对这一“自然实验”和一名表型相似的青春期女性的研究，使我们发现了一种在包括男性在内的灵长类动物中抵抗维生素D和雌激素的新方法。我们创造了这些导致抗性的蛋白质，维生素D反应元件结合蛋白(VDRE-BP)和雌激素反应元件结合蛋白(ERE-BP)，因为它们能够与连接的维生素D受体(VDR)和雌激素受体1(ER1)竞争其同源反应元件的反式，确定它们是异质性核糖核蛋白C(HNRNPC)家族的核酸结合蛋白。最近的工作使我们认识到，这些hnRNP改变类固醇激素导向的基因表达的能力并不局限于它们在转录水平上的显性负作用。这些hnRNP具有与单链DNA(SsDNA)、单链DNA(SsRNA)和双链DNA(DsDNA)相互作用的能力，因此有可能控制细胞内多个位置的基因表达。在这里，我们的理论是，特定的hnRNP可以作为单个基因产物同步表达的多位点参与者，依次调节染色质重塑(ssDNA结合)、转录(dsDNA结合)、剪接(ssDNA和ssRNA结合)和microRNA(miRNA；ssRNA结合)。由于我们所研究的人类和亚人类灵长类动物对hnRNP过度表达的自然界实验都导致了对青春期女性生长骨骼的维生素D和雌激素拮抗作用，因此这里我们将集中讨论这些hnRNP的自然靶点--生长的骨骼和成骨细胞。该研究提出了三个具体目标：1)利用染色质修饰/沉淀、过低表达和新颖的细胞自由翻译技术，探讨hnRNPs对负责1，25-二羟基维生素D和雌二醇驱动的骨生长的细胞的影响；2)采用新型RNA免疫沉淀和Micro-RNA方法研究这些hnRNPs的通用性，通过定向结合同一基因的RNA产物来控制转录剪接和处理，该产物的表达同时在染色质重塑和转录水平上受到调节；以及3)通过靶向在小鼠骨中过表达VDBP RE-BP和ere-BP hnRNPs来确定这些hnRNPs对生长中的青少年骨骼的综合影响。我们期望这项拟议的研究将为从机制上理解这些多功能hnRNP如何控制生长骨中类固醇/类固醇激素调节基因表达的过程铺平道路，并勾勒出在这一过程中潜在的治疗干预的新观点。与公共健康相关：维生素D和雌激素是两种类固醇荷尔蒙，长期以来一直被认为能够影响骨骼的生长。25年前，我们开始调查居住在洛杉矶动物园的青少年、雌性新大陆灵长类动物中的软骨病暴发，鉴定了异质核核糖核蛋白C(HNRNPC)家族的蛋白质、维生素D反应元件结合蛋白(VDRE-BP)和雌激素反应元件结合蛋白(ERE-BP)，这些蛋白质导致对维生素D和雌激素对骨骼的作用产生抵抗。凭借它们与单链DNA(SsDNA)、ssRNA以及双链DNA(DsDNA)相互作用的能力，我们计划进行实验来验证我们的理论，即VDRE-BP和ERE-BP可以作为单个基因产物的同步表达的多位点参与者在骨骼中发挥作用，从而依次调节i)染色质重塑，ii)转录，iii)转录剪接和iv)转录处理。

项目成果

期刊论文数量（26）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Vitamin D insufficiency and skeletal development in utero.

DOI：
10.1002/jbmr.2
发表时间：
2010-01
期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
0
作者：
Hewison M;Adams JS
通讯作者：
Adams JS

Vitamin D and alternative splicing of RNA.

维生素 D 和 RNA 的选择性剪接。

DOI：
10.1016/j.jsbmb.2014.09.025
发表时间：
2015
期刊：
The Journal of steroid biochemistry and molecular biology
影响因子：
0
作者：
Zhou,Rui;Chun,ReneF;Lisse,ThomasS;Garcia,AlejandroJ;Xu,Jianzhong;Adams,JohnS;Hewison,Martin
通讯作者：
Hewison,Martin

Endogenous blockade of 1,25-dihydroxyvitamin D-receptor binding in New World primate cells.

新大陆灵长类细胞中 1,25-二羟基维生素 D 受体结合的内源性阻断。