LIM kinase 1 and Cell Cycle Regulation

LIM 激酶 1 和细胞周期调节

基本信息

批准号：
7522364
负责人：
RATNA CHAKRABARTI
金额：
$ 21.3万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7522364
关键词：
Actins Activator Appliances Aneuploidy Area BCL-2 Protein BCL2 gene Binding Biological Assay Cell Cycle Progression Cell Cycle Regulation Cell Line Cell Proliferation Cell Shape Cell physiology Cells Centrosome Chromosomal Instability Chromosome abnormality Complementary DNA Complex Cyclin D1 Cyclin E Cyclin-Dependent Kinase Inhibitor Cyclin-Dependent Kinases Cyclins Cytokinesis Cytoskeleton Data Defect Development Diagnosis Disease Ectopic Expression Environment Epithelial Cells Equilibrium Event Exhibits Fibroblasts Funding G1 Arrest G1 Phase Genomic Instability Gills Goals Growth Growth Factor Hormones Human Individual Knowledge LIM Domain Kinase 1 Laboratories Lead Learning Literature MSLN gene Maintenance Malignant Neoplasms Microtubule Polymerization Microtubules Mission Mitosis Mitotic Mitotic spindle Normal Cell Nuclear Outcome Pathogenesis Pathologic Pharmaceutical Preparations Phase Phase Transition Phosphorylation Phosphotransferases Play Preventive Process Protein Dephosphorylation Protein Overexpression Protein-Serine-Threonine Kinases Proteins Public Health Purpose Qualifying Reagent Refractory Regulation Research Research Personnel Resistance Resource Sharing Role Seminal Series Signal Pathway Signal Transduction Site Structure Study Section Testing Therapeutic Therapeutic Intervention Time Translating Variant Work Yang actin depolymerizing proteins base cancer cell cancer type cell growth regulation cofilin cyclin G1 depolymerization experience genetic regulatory protein inhibitor/antagonist innovation mutant novel polymerization protein activation protein function protein protein interaction response stable cell line tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): There is a gap in understanding how variation in expression of cytoskeleton modulatory proteins leads to abnormal progression of cell cycle resulting in aneuploidy, centrosomal multiplication and altered cell proliferation. The long-term goal is to understand better the underlying mechanism of individual steps of G1/S phase progression and how LIM kinase 1, an actin and microtubule cytoskeleton modulatory protein, is involved in G1 to S phase progression as variation in expression of LIM kinase 1 induced transient G1/S arrest, delayed progression of G2/M and centrosome multiplication. The objective of this particular application is to identify how initial events of G1 progression such as expression of early G1 regulatory proteins, activation of G1 Cyclin/Cdks and inactivation of Cdk inhibitors is regulated through protein phosphorylation and dephosphorylation, and how LIM kinase 1 participates in these processes; also, to determine how variation in expression of LIMK1 leads to sluggish progression of G1 cells to the S phase. The central hypothesis is that LIMK1 negatively regulates progression of cells through G1 to S phase through altered localization and functions of activators and inhibitors of G1 and S phase Cdks. The rationale for the proposed research is that understanding the fundamental mechanism of G1/S phase progression has the potential to translate into better understanding the pathogenesis of abnormal expression of LIMK1 in epithelial cell cancers. Thus, the proposed research is relevant to that part of NIH's mission that pertains to developing fundamental knowledge that will potentially help to reduce the burdens of human cancers. Guided by strong preliminary data, this hypothesis will be tested by two specific aims: 1) Examine the expression, localization of G1/S phase regulatory proteins and activation status of LIMK1 in G0 synchronized RWPE1 cells expressing LIMK1 in response to growth factor stimulation and 2) Elucidate interactions of G1/S phase regulatory proteins through complex formation leading to functional activation or inactivation of Cyclin/Cdks in response to mitogenic signals. Under specific aim 1 we will use the approach of overexpression of LIMK1 and utilize stable cell lines to determine specific effects of expression, phosphorylation and catalytic function of LIMK1 on progression of G1 phase, and expression of G1 and S phase kinases and their inhibitors. Under the second aim, activation of G1 and S phase Cdks and stability of their activator and inhibitors will be examined using kinase assays. This approach is innovative because it capitalizes on a new observation from our group to obtain additional information on the mechanism of G1 to S phase transition. The proposed research is significant, because it is expected to advance and expand the understanding of how kinases influence G1/S phase progression and how aberrant expression of these kinases interferes with normal cell cycle progression. PUBLIC HEALTH RELEVANCE: The proposed studies are of an important and under investigated area of regulation of G1/S phase and function of LIM kinase 1 that has potential applicability to understanding the pathogenesis caused by abnormal expression of LIM kinase 1 in cancer. LIM kinase 1 regulates cellular functions through protein-protein interaction and phosphorylation, and maintains cell shape through remodeling of actin cytoskeleton. The proposed research has relevance to public health, because it investigates a fundamental mechanism of cell cycle progression, which often becomes aberrant during development of cancer.

描述（由申请人提供）：对于细胞骨架调节蛋白表达的变化如何导致细胞周期异常进展导致非整倍性、中心体增殖和细胞增殖改变的理解存在差距。长期目标是更好地了解 G1/S 期进展各个步骤的潜在机制，以及 LIM 激酶 1（一种肌动蛋白和微管细胞骨架调节蛋白）如何通过 LIM 激酶表达的变化参与 G1 至 S 期进展1 诱导短暂的 G1/S 停滞，延迟 G2/M 的进展和中心体增殖。该特定应用的目的是确定 G1 进展的初始事件（例如早期 G1 调节蛋白的表达、G1 细胞周期蛋白/Cdks 的激活和 Cdk 抑制剂的失活）如何通过蛋白磷酸化和去磷酸化进行调节，以及 LIM 激酶 1 如何参与这些过程；此外，还确定 LIMK1 表达的变化如何导致 G1 细胞缓慢进展至 S 期。核心假设是，LIMK1 通过改变 G1 和 S 期 Cdks 激活剂和抑制剂的定位和功能，负向调节细胞从 G1 到 S 期的进展。这项研究的基本原理是，了解 G1/S 期进展的基本机制有可能转化为更好地了解上皮细胞癌中 LIMK1 异常表达的发病机制。因此，拟议的研究与 NIH 使命的一部分相关，即开发可能有助于减轻人类癌症负担的基础知识。在强有力的初步数据指导下，该假设将通过两个具体目标进行检验：1）检查表达 LIMK1 的 G0 同步 RWPE1 细胞响应生长因子刺激的 G1/S 期调节蛋白的表达、定位和 LIMK1 的激活状态，2） ) 阐明 G1/S 期调节蛋白通过复合物形成的相互作用，导致 Cyclin/Cdks 响应有丝分裂信号的功能激活或失活。在具体目标1下，我们将使用LIMK1过表达的方法并利用稳定细胞系来确定LIMK1的表达、磷酸化和催化功能对G1期进展以及G1和S期激酶及其抑制剂表达的具体影响。第二个目标是使用激酶测定来检查 G1 和 S 期 Cdks 的激活及其激活剂和抑制剂的稳定性。这种方法是创新的，因为它利用我们小组的新观察来获取有关 G1 到 S 相变机制的更多信息。拟议的研究意义重大，因为它有望推进和扩大对激酶如何影响 G1/S 期进程以及这些激酶的异常表达如何干扰正常细胞周期进程的理解。公共健康相关性：拟议的研究属于 LIM 激酶 1 G1/S 期和功能调节的重要且正在研究的领域，对于了解癌症中 LIM 激酶 1 异常表达引起的发病机制具有潜在的适用性。 LIM 激酶 1 通过蛋白质-蛋白质相互作用和磷酸化调节细胞功能，并通过肌动蛋白细胞骨架重塑维持细胞形状。拟议的研究与公共健康相关，因为它研究了细胞周期进展的基本机制，细胞周期进展在癌症发展过程中经常变得异常。