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）：了解细胞骨架调节蛋白表达的变化如何导致细胞周期的异常进展，从而导致非整倍性，中心体繁殖和细胞增殖改变。长期目标是更好地了解G1/S相进展的各个步骤的潜在机制，以及LiM激酶1（肌动蛋白和微管细胞骨架调节蛋白）如何参与G1与S相位的S阶段，因为LIM激酶1在LIM激酶1的表达变化中诱导的瞬时G1/S s抑制G1/s抑制，G2/M延迟，延迟，延迟，延迟，延迟，延迟。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/CDK的激活以及CDK抑制剂的灭活如何通过蛋白质磷酸化和去磷酸化以及LIM激酶1在这些过程中参与这些过程；同样，为了确定LIMK1表达的变化如何导致G1细胞向S相的进展缓慢。中心假设是，Limk1通过改变了G1和S相CDK的活化剂和抑制剂的定位和功能，对细胞通过G1到S相进行负面调节。拟议的研究的基本原理是，了解G1/S相进展的基本机制具有更好地理解上皮细胞癌中Limk1异常表达的发病机理的潜力。因此，拟议的研究与NIH使命的那部分有关，该研究与发展基本知识有关，这有可能有助于减轻人类癌症的负担。在强有力的初步数据的指导下，该假设将通过两个具体目的检验：1）检查G1/s相调节蛋白的表达，定位，G1/s相位调节蛋白的定位和LIMK1在G0同步的RWPE1细胞中表达LIMK1的LIMK1的激活状态，对生长因子刺激的响应以及2）阐明G1/S相互调节蛋白的相互作用的相互作用或2）响应有丝分裂信号的细胞周期蛋白/CDK。在特定目标1下，我们将使用Limk1过度表达的方法，并利用稳定的细胞系来确定Limk1表达，磷酸化和催化功能对G1相进展的特定作用，G1和S相激酶及其抑制剂的表达。在第二个目标下，将使用激酶测定法检查G1和S期CDK的激活以及其活化剂和抑制剂的稳定性。这种方法具有创新性，因为它利用了我们小组的新观察结果，以获取有关G1到S相变机理的其他信息。拟议的研究很重要，因为它有望提高和扩展对激酶如何影响G1/s相位进展的理解以及这些激酶的异常表达如何干扰正常的细胞周期进程。公共卫生相关性：拟议的研究是一个重要的，并且在LIM激酶1的G1/S相调节和功能的调节区域中具有潜在的适用性，可在理解癌症中LIM激酶1异常表达引起的发病机理。 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，并通过重塑肌动蛋白细胞骨架来维持细胞形状。拟议的研究与公共卫生有关，因为它研究了细胞周期进展的基本机制，在癌症的发展过程中通常会变得异常。