Role of JAK2-PAK1 interaction in prolactin-dependent signaling

JAK2-PAK1 相互作用在催乳素依赖性信号传导中的作用

基本信息

批准号：
8136055
负责人：
MARIA DIAKONOVA
金额：
$ 29.89万
依托单位：
UNIVERSITY OF TOLEDO
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：
2010-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8136055
关键词：
Actins Adipocytes Affect Alveolar Anterior Pituitary Gland Apoptosis Astrocytes Basic Science Binding Biological Biological Process Breast Breast Cancer Cell CCND1 gene Cancer Center Cancer Research Project Cancer cell line Cell Survival Cells Cellular Morphology Cessation of life Clinical Sciences Columbidae Consult Cyclin D1 Cytokine Receptors Cytoskeleton DNA Sequence Rearrangement Data Development Dissection Endocrine Epithelial Cells Etiology Event Functional disorder Gene Expression Regulation Genes Genetic Transcription Gland Goals Grant Growth Factor Hormones Human JAK2 gene Lacrimal gland structure Lactation Letters Life Expectancy Ligand Binding Link Liver Location Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Mammary Neoplasms Mammary gland Mediating Membrane Michigan Milk Molecular Molecular Target Multiprotein Complexes Names Normal Cell Oryctolagus cuniculus Ovary Pancreas Pathway interactions Peptide Mapping Pharmaceutical Preparations Phosphorylation Phosphotransferases Physician Executives Pituitary Gland Play Production Prolactin Prolactin Receptor Prostate Protein Tyrosine Kinase Protein-Serine-Threonine Kinases Proteins Receptor Activation Receptor Signaling Recruitment Activity Regulation Research Risk Role Signal Pathway Signal Transduction Signaling Molecule Signaling Protein Specimen Structure of beta Cell of islet Submandibular gland T-Lymphocyte Translational Research Tumorigenicity Tyrosine Tyrosine Phosphorylation United States Universities Woman Work adapter protein autocrine base biological adaptation to stress cancer cell cancer diagnosis cancer type carcinogenesis cell motility cell type design human disease in vivo interest malignant breast neoplasm mammary epithelium mammary gland development medical schools neoplastic cell novel novel therapeutic intervention oncology overexpression protein protein interaction public health relevance receptor receptor binding response tumor tumor progression two-dimensional

项目摘要

DESCRIPTION (provided by applicant): In normal mammary development, the hormone prolactin (PRL) is critical for alveolar proliferation and differentiation. Increasing evidence supports the involvement of PRL in breast cancer, the leading type of cancer in women and the second leading cause (after lung cancer) of cancer death among women. In 2008, 40,480 women are expected to die from breast cancer in the U.S. The prolactin receptor (PRLR) is detected in 80% of human breast cancers and is overexpressed in breast cancer cells. Normal and tumor mammary epithelial cells synthesize PRL and PRLR, thus the PRL could behave as an autocrine growth factor for human breast cancer cells. These results suggest the need for a more complete understanding of PRLR signaling in breast cancer. Tyrosine (Tyr) kinase JAK2 was identified as a PRLR-bound signaling molecule. Identification of the proteins recruited to the PRLR-JAK2 and dissection of the signaling pathways that are subsequently activated will ultimately provide a basis for understanding PRL action. Preliminary data demonstrate that the serine-threonine kinase PAK1 associates with and is Tyr phosphorylated by JAK2. Two-dimensional peptide mapping identified three Tyr(s) of PAK1 which are phosphorylated by JAK2. Tyr phosphorylation of PAK1 by JAK2 was also shown to increase cell motility. In this grant we propose to examine the hypothesis prolactin-dependent JAK2 phosphorylation of PAK1 regulates PAK1 activity. Activated PAK1 regulation of target proteins may depend on phosphorylation events or/and protein-protein interactions, leading to the formation of a multiprotein complex that modulates the actin cytoskeleton, increases cell motility and invasiveness, mediates cyclin D1 gene transcription and affects tumorigenicity of human breast cancer cells. Aim1 will determine the role of JAK2-phosphorylated PAK1 in regulating PRL-dependent actin cytoskeleton rearrangement, cell motility and invasiveness. Aim2 will determine the role of PAK1 in regulating PRL-activated cyclin D1 gene transcription. Finally, Aim3 will determine whether JAK2 phosphorylation of PAK1 affects the tumorigenicity of human breast cancer cells in vivo. Because both PAK1 and PRL have been implicated in breast cancer, the proposed studies may ultimately fill out the existing gap between upstream PRL-PRLR-JAK2 events and downstream PAK1-dependent functions in our understanding of the mechanism of human breast cancer. Tyr phosphorylation of PAK1 by JAK2 is likely to represent a novel molecular target in the search for the etiology and treatment of human breast cancer. PUBLIC HEALTH RELEVANCE: Prolactin (PRL) was discovered in 1928 as a pituitary factor able to stimulate mammary gland development and lactation in rabbits, as well as the production of crop milk in pigeons. A few years later, the name prolactin was given, based on its ability to stimulate milk production. More than 300 separate biological activities have been attributed to PRL. These biological functions are mediated by specific membrane receptors. These receptors are non tyrosine kinases, they transduce the signal via associated kinases that are recruited by the receptor and activated upon ligand binding. One of these kinases is JAK2 tyrosine kinase. We have recently shown that another protein - PAK1 - is a novel substrate of JAK2. We showed that JAK2 binds to PAK1 and makes PAK1 more active. Activated PAK1 contributes to better cell survival and cell migration. However, how these two proteins work together and the precise mechanism of their action is unknown. PRL is also involved in breast cancer. Breast cancer is the most commonly diagnosed cancer in women - nearly 1 in 3 (30%) of all cancers in women occur in the breast. Based on the current life expectancy for women in the United States, 1 out of 9 women will develop breast cancer in her lifetime - a risk that was 1 out of 14 in 1960. Much is still unknown regarding the molecular biological mechanism by which a normal cell becomes a cancer cell. Our long term goal is to understand the molecular mechanisms of PRL action and disregulation of which leads to human diseases including breast cancer. Toward this aim, we have started to analyze the relationship between JAK2 and PAK1. In the current proposal we will study how JAK2 and PAK1 increase cell migration and cell survival, and which genes are regulated by these two proteins. Understanding how these proteins work together will help to design new therapeutic approaches and possibly drugs for treatment of different human diseases and breast cancer.

描述（由申请人提供）：在正常的乳腺发育中，激素催乳素（PRL）对于肺泡增殖和分化至关重要。越来越多的证据支持PRL参与乳腺癌，女性的主要癌症类型，以及妇女中癌症死亡的第二大主要原因（肺癌之后）。在2008年，预计在美国，有40,480名女性死于乳腺癌，在80％的人类乳腺癌中检测到催乳素受体（PRLR），并且在乳腺癌细胞中过表达。正常和肿瘤乳腺上皮细胞合成PRL和PRLR，因此PRL可以作为人类乳腺癌细胞的自分泌生长因子。这些结果表明需要对乳腺癌中的PRLR信号传导有更全面的了解。酪氨酸（Tyr）激酶JAK2被鉴定为PRLR结合的信号分子。鉴定募集到PRLR-JAK2的蛋白质和随后激活的信号通路的解剖最终将为理解PRL作用提供基础。初步数据表明，丝氨酸 - 硫代激酶PAK1与JAK2磷酸化并与Tyr磷酸化。二维肽映射确定了三个由JAK2磷酸化的PAK1的Tyr。 JAK2对PAK1的Tyr磷酸化也显示出增加细胞运动性。在这笔赠款中，我们建议检查PAK1的假设催乳素依赖性JAK2磷酸化调节PAK1活性。靶蛋白的活化PAK1调节可能取决于磷酸化事件或/和蛋白质 - 蛋白质相互作用，从而导致形成多动蛋白复合物，从而调节肌动蛋白细胞骨架，增加细胞的运动和侵入性，介导细胞周期蛋白D1基因转录并影响人乳腺癌细胞的肿瘤性肿瘤性。 AIM1将确定JAK2磷酸化PAK1在调节PRL依赖性肌动蛋白细胞骨架重排，细胞运动和侵入性中的作用。 AIM2将确定PAK1在调节PRL激活的细胞周期蛋白D1基因转录中的作用。最后，AIM3将确定PAK1的JAK2磷酸化是否会影响体内人类乳腺癌细胞的肿瘤性。由于PAK1和PRL都与乳腺癌有关，因此提出的研究最终可能填补了上游PRL-PRLR-JAK2事件与下游PAK1依赖性功能之间现有的差距，这在我们对人类乳腺癌机制的理解中的理解。 JAK2对PAK1的Tyr磷酸化可能代表着寻找人类乳腺癌病因和治疗的新分子靶标。公共卫生相关性：催乳素（PRL）于1928年被发现是一种垂体因素，能够刺激兔子的乳腺发育和泌乳，以及鸽子的作物牛奶的产生。几年后，基于其刺激牛奶产量的能力，给出了催乳素的名称。超过300种独立的生物活动归因于PRL。这些生物学功能是由特定的膜受体介导的。这些受体是非酪氨酸激酶，它们通过受体募集并在配体结合时激活的相关激酶传递信号。这些激酶之一是JAK2酪氨酸激酶。我们最近表明，另一种蛋白质-PAK1-是JAK2的新型底物。我们表明JAK2与PAK1结合，并使PAK1更活跃。活化的PAK1有助于更好的细胞存活和细胞迁移。但是，这两种蛋白质如何一起起作用以及其作用的确切机制尚不清楚。 PRL也参与乳腺癌。乳腺癌是女性最常见的癌症 - 乳房中近三分之一（30％）的癌症发生。基于当前美国女性的预期寿命，在她的一生中，有1名女性将患上乳腺癌 - 1960年14岁的风险在1960年中有1个。对于正常细胞成为癌细胞的分子生物学机制，仍然未知很多。我们的长期目标是了解PRL作用的分子机制和无调导致包括乳腺癌在内的人类疾病。为了实现这一目标，我们已经开始分析JAK2和PAK1之间的关系。在当前的提案中，我们将研究JAK2和PAK1如何增加细胞迁移和细胞存活，哪种基因受这两种蛋白质的调节。了解这些蛋白质如何共同起作用将有助于设计新的治疗方法，并可能使用用于治疗不同人类疾病和乳腺癌的药物。