Developmental & Molecular Mechanisms Employed by Chato, KRAB Zinc Finger Protein

发育性

基本信息

批准号：
8468722
负责人：
Maria J Garcia-Garcia
金额：
$ 27.34万
依托单位：
CORNELL UNIVERSITY
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：
2010-09-15 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8468722
关键词：
Affect Amphibia Anterior Anxiety Behavior Binding Biological Assay Boxing Cells Characteristics Chimera organism Chromatin Complex Congenital Abnormality DNA DNA Sequence DNA Sequence Rearrangement Defect Development Developmental Gene Disease Early Diagnosis Embryo Embryonic Development Endoderm Enzymes Event Family member Gene Family Gene Targeting Genetic Genetic Screening Genetic Transcription Goals Human Individual Knowledge Lead Mammals Mediating Membrane Molecular Morphogenesis Mus Mutation Neoplasm Metastasis Neural Tube Defects Neural tube Phenotype Physiological Processes Pregnant Women Prevention Process Proteins Recruitment Activity Regulation Research Research Project Grants Risk Factors Role Shapes Signal Pathway Signal Transduction Somites Specificity Spinal Dysraphism Staging TRIM Motif Testing Tissues Transcription Repressor/Corepressor Vertebrates Wild Type Mouse Xenopus Yeasts Yolk Sac Zebrafish Zinc Fingers base cell motility chromatin immunoprecipitation embryo tissue imprint in vivo insight interest malformation mammalian genome mutant novel polarized cell positional cloning public health relevance research study stem cell differentiation teleost tool yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): The goal of the proposed research project is to understand the developmental and molecular mechanisms that control morphogenesis during mammalian development. Our research is focused on the study of convergent extension, a process that mediates elongation of tissues and involves highly coordinated rearrangements/movements of cells. Numerous devastating congenital malformations result from aberrant coordination of cell movements, but the genetic basis of these diseases is in most cases unknown, which impedes their early diagnosis and prevention. Studies in amphibian and teleost vertebrates have revealed that non-canonical Wnt signaling regulates convergent extension. However, studies in mouse have indicated that convergent extension might be regulated by additional molecular mechanisms in mammals. A phenotype-based forward genetic screen identified chato, a mouse mutation causing embryonic lethality and convergent extension phenotypes. Our analysis of chato mutants confirmed that chato is required for the proper rearrangement of cells that lead to definitive endoderm convergent extension. chato encodes ZFP568, a novel Kruppel-Associated-Box (KRAB) Zinc finger protein. Analysis of chato embryos suggests that chato does not implement non-canonical Wnt signaling, but rather controls mouse elongation through an independent molecular mechanism. One of the goals of this proposal is to identify the molecular effectors and signaling pathways employed by chato to control convergent extension. We are also interested in understanding the developmental mechanisms utilized by chato: In addition to their convergent extension defects, chato mutants also display malformations in morphogenesis of extraembryonic tissues, which might indicate a role for these supporting tissues in promoting convergent extension. We will perform experiments to test this hypothesis. We have divided our research plan into three specific aims: In Specific Aim 1, we will determine whether extraembryonic tissues influence convergent extension of the embryo by analyzing the phenotype of tetraploid chimeras and genetic mosaic embryos. The study of chatwo mutants has provided genetic evidence supporting a role for TRIM28 as a transcriptional co-repressor of ZFP568. In Specific Aim 2, we determine the factors that influence formation of ZFP568-TRIM28 complexes and the mechanisms that mediate the specificity of this complex to the control of convergent extension. Finally, in Specific Aim 3, we seek to identify direct and indirect targets of ZFP568 and ZFP568-TRIM28 complexes and determine the ultimate ZFP568 effectors that control convergent extension. Overall, results from our experiments will increase our understanding of the molecules that control mammalian morphogenesis and provide insights about the molecular mechanisms utilized by KRAB Zinc finger proteins.

描述（由申请人提供）：拟议的研究项目的目标是了解控制哺乳动物发育过程中形态发生的发育和分子机制。我们的研究集中在收敛扩展的研究上，该过程介导组织的伸长并涉及高度协调的重排/运动。许多毁灭性的先天性畸形是由于细胞运动的异常协调而导致的，但是这些疾病的遗传基础在大多数情况下是未知的，这阻碍了它们的早期诊断和预防。在两栖动物和硬骨脊椎动物中的研究表明，非典型的Wnt信号传导调节收敛扩展。然而，小鼠的研究表明，收敛扩展可能受哺乳动物中的其他分子机制调节。基于表型的正向遗传筛选确定了Chato，这是一种小鼠突变，导致胚胎致死性和收敛性延伸表型。我们对Chato突变体的分析证实，Chato对于导致确定性内胚层收敛扩展的细胞的适当重排是必需的。 Chato编码ZFP568，这是一种新型的Kruppel相关盒（KRAB）锌指蛋白。对Chato胚胎的分析表明，Chato不会实现非典型的Wnt信号传导，而是通过独立的分子机制来控制小鼠的伸长。该提案的目标之一是确定Chato采用的分子效应子和信号传导途径来控制收敛扩展。我们还有兴趣了解Chato所使用的发展机制：除了它们的收敛扩展缺陷外，Chato突变体还显示出畸形的畸形外胚膜组织的形态发生，这可能表明这些支持组织在促进收敛扩展中的作用。我们将执行实验来检验这一假设。我们将研究计划分为三个特定目标：在特定的目标1中，我们将通过分析四倍体嵌合体和遗传镶嵌胚胎的表型来确定胚外组织是否会影响胚胎的收敛扩展。 Chatwo突变体的研究提供了支持TRIM28作为ZFP568的转录共抑制剂的遗传证据。在特定目标2中，我们确定了影响ZFP568-TRIM28复合物形成的因素，以及介导该复合物对收敛扩展的特异性的机制。最后，在特定目标3中，我们试图确定ZFP568和ZFP568-TRIM28复合物的直接和间接目标，并确定控制收敛扩展的最终ZFP568效应子。总体而言，我们的实验结果将增加我们对控制哺乳动物形态发生的分子的理解，并提供有关KRAB锌指蛋白使用的分子机制的见解。