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）：通过调节核和细胞质定位来控制转录因子活性是一种重要且进化保守的机制。配体调节转录因子的核受体（NR）的成员经历了核进出口周期。我们发现，前列腺癌细胞中雄激素受体（AR）的核输出受磷酸化调节。经历雄激素诱导的磷酸化的七个部位中两个中两个的丙氨酸突变导致AR导出到细胞质的缺陷。一个调节AR出口的部位（SER308）在N末端域中。另一个位点（SER650）位于AR的铰链区域，靠近DMA结合域，该结构域是核输出信号的功能。我们生成了一系列AR磷材料特异性抗体，发现Ser650在体外和体内被应力激活的MAP激酶P38磷酸化。我们发现高渗透诱导了从细胞核到细胞质的AR重新分布。在AR磷酸化位点，AR重新分布可通过丙氨酸突变阻断，不需要新的蛋白质合成或导出受体CRM1，并且是可逆的。该提议的假设是，环境应力激活的多个信号转导途径调节核质转运的过程。在目标1和2中，我们将确定调节AR重新分布的应力激酶途径（P38，JNK，ERK），确定AR重新分布是否是由于刺激核出口或进口的抑制，并确定使用抑制剂，siRNA和敲除细胞来确定单个激酶的贡献。除了调节AR的定位外，应力信号传导通过破坏RAN梯度对大多数核运输途径至关重要的RAN梯度发挥全球影响。在AIM 3中，我们将确定RAN梯度破坏是否涉及控制运输转运或RAN GTPase周期的因素。我们的研究将为介导信号转导，核转运和NR活性之间的串扰的机制提供新的见解。

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.

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。