权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Pathways of Nucleocytoplasmic Transport

核细胞质运输途径

基本信息

批准号：
7935045
负责人：
Bryce Paschal
金额：
$ 11.56万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7935045
关键词：
Address Affect Alanine Androgen Antagonists Androgen Receptor Androgens Antibodies Binding Biology Cell Nucleus Cells Cellular biology Chemicals Coculture Techniques Collaborations Cytoplasm Defect Diagnostic Neoplasm Staging Disease Eukaryota Family Genes Global Change Growth Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Hydrolysis In Vitro Individual Knock-out LNCaP Laboratories Ligand Binding Ligands MAPK14 gene MAPK8 gene Malignant neoplasm of prostate Mammals Mass Spectrum Analysis Massachusetts Mediating Mitogen-Activated Protein Kinases Modeling Molecular Mutation N-terminal Nuclear Nuclear Export Nuclear Import Nuclear Receptors Nucleotides PC3 cell line Pathway interactions Patients Phosphorylation Phosphorylation Site Phosphotransferases Plasmids Play Post-Translational Protein Processing Process Prostate Prostate Cancer therapy Protein Biosynthesis Proteins Reagent Regulation Research Personnel Role Running Signal Transduction Signal Transduction Pathway Site Small Interfering RNA Stress System Testing Tumor stage base bone cell cancer cell hormone therapy in vivo inhibitor/antagonist insight intercellular communication member mitogen-activated protein kinase p38 nuclear transport factor 2 nucleocytoplasmic transport protein complex receptor receptor function response transcription factor upstream kinase

项目摘要

DESCRIPTION (provided by applicant): Controlling transcription factor activity by regulating nuclear and cytoplasmic localization is an important and evolutionarily conserved mechanism. Members of the nuclear receptor (NR) super-family of ligand regulated transcription factors undergo cycles of nuclear import and export. We have found that nuclear export of the androgen receptor (AR) in prostate cancer cells is regulated by phosphorylation. Alanine mutations in two of the seven sites that undergo androgen-induced phosphorylation cause a defect in AR export to the cytoplasm. One site that regulates AR export (Ser308) is in the N-terminal domain. The other site (Ser650) is in the hinge region of AR, proximal to the DMA binding domain that functions as a nuclear export signal. We generated a panel of AR phosphosite-specific antibodies and found that Ser650 is phosphorylated by the stress-activated MAP kinase p38 in vitro and in vivo. We found that hyperosmolarity induces AR redistribution from the nucleus to the cytoplasm. This AR redistribution to the cytoplasm can be blocked by alanine mutations in AR phosphorylation sites, does not require new protein synthesis or the export receptor Crm1, and is reversible. The hypothesis of this proposal is that multiple signal transduction pathways activated by environmental stress regulate the process of nucleocytoplasmic transport. In Aims 1 and 2 we will identify the stress kinase pathways (p38, JNK, ERK) that regulate AR redistribution, determine if the AR redistribution is due to stimulation of nuclear export or inhibition of import, and determine the contributions of individual kinases using inhibitors, siRNA, and knockout cells. In addition to regulating the localization of AR, stress signaling exerts global effects on the nuclear transport machinery by disrupting the Ran gradient which is essential for most nuclear transport pathways. In Aim 3 we will determine whether Ran gradient disruption involves stress signaling to factors that control Ran transport or the Ran GTPase cycle. Our studies will provide new insight into the mechanisms that mediate crosstalk between signal transduction, nuclear transport, and NR activity.

描述(由申请人提供)：通过调节细胞核和细胞质的定位来控制转录因子的活性是一个重要的和进化上保守的机制。核受体(NR)超家族的配体调节转录因子经历了核输入和输出周期。我们发现前列腺癌细胞中雄激素受体(AR)的核输出受磷酸化的调节。在经历雄激素诱导的磷酸化的七个位置中，有两个位置的丙氨酸突变导致AR输出到细胞质的缺陷。调节AR输出的一个站点(Ser308)位于N端域。另一个位点(Ser650)位于AR的铰链区，靠近作为核输出信号的DMA结合域。我们产生了一组AR亚磷酸盐特异性抗体，并在体外和体内发现Ser650被应激激活的MAP激酶p38磷酸化。我们发现，高渗诱导AR从胞核到胞浆的重新分布。这种AR到细胞质的重新分布可以被AR磷酸化位点的丙氨酸突变所阻断，不需要新的蛋白质合成或出口受体CRM1，并且是可逆的。这一设想的假设是，环境应激激活的多条信号转导通路调节核质运输过程。在AIMS 1和AIMS 2中，我们将确定调节AR再分配的应激激酶通路(p38、JNK、ERK)，确定AR再分配是由于核输出的刺激还是进口的抑制，并确定使用抑制剂、siRNA和基因敲除细胞的单个激酶的贡献。除了调节AR的定位外，应激信号还通过扰乱RAN梯度对核运输机制产生全球影响，RAN梯度是大多数核运输途径所必需的。在目标3中，我们将确定RAN梯度中断是否涉及向控制RAN运输或RAN GTP酶循环的因素发出压力信号。我们的研究将为调节信号转导、核运输和NR活性之间的串扰机制提供新的见解。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

NXT1 is necessary for the terminal step of Crm1-mediated nuclear export.

NXT1对于CRM1介导的核出口的终端步骤是必需的。

DOI：
10.1083/jcb.152.1.141
发表时间：
2001-01-08
期刊：
The Journal of cell biology
影响因子：
0
作者：
Black BE;Holaska JM;Lévesque L;Ossareh-Nazari B;Gwizdek C;Dargemont C;Paschal BM
通讯作者：
Paschal BM

Scientists share nuclear secrets at Jekyll Island.

科学家们分享杰基尔岛的核秘密。