Developmental analysis of SELCT proteins

SELCT蛋白的发育分析

• 批准号: 7629768
• 负责人: Anne Church Hart
• 金额: $ 8.04万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2009-11-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7629768
• 关键词: Address; Alagille Syndrome; Alleles; Alzheimer' s Disease; Amino Acid Motifs; Binding; Biochemistry; Biological Assay; CADASIL; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Lineage; Cell Maintenance; Conserved Sequence; Defect; Dementia; Development; Disease; Drosophila genus; Dysostoses; EGF gene; Family; Genes; Genetic; Genetic Epistasis; Genome; Homologous Gene; Immunohistochemistry; In Vitro; Inborn Genetic Diseases; Invertebrates; Lateral; Ligands; Malignant Neoplasms; Membrane; Memory; Modeling; Molecular; Molecular Genetics; Mutation; Names; Notch Signaling Pathway; Pathology; Pathway interactions; Pattern; Phenotype; Play; Process; Proteins; RNA Interference; Receptor Activation; Regulation; Research Personnel; Role; Signal Transduction; Stem cells; Stroke; Structure; Testing; Transgenic Organisms; Variant; Vertebrates; base; cell fate specification; effective therapy; extracellular; genetic analysis; glycosylation; in vitro testing; in vivo; mutant; notch protein; novel; presenilin; programs; receptor; receptor binding; research study; therapy development; tissue culture; yeast two hybrid system

DESCRIPTION (provided by applicant): The conserved Notch signaling pathway is critical for cell fate specification and lateral inhibition during development in vertebrates and invertebrates. Notch receptors bind extracellular ligands (e.g. Delta, Serrate and LAG-2) containing a conserved DSL domain that is critical for receptor activation. We have identified a new family of putative Notch ligands in C. elegans with homologs in vertebrates. Unlike canonical Notch ligands, these proteins lack the conserved DSL domain but they share a conserved sequence motif with classical Notch ligands. The first aim of this proposal examines in detail the function of one of these new putative ligands. SEL-14 is required for normal development and preliminary studies suggest that it activates Notch signaling. The role of SEL-14 in Notch signaling will be examined at a molecular and cellular level using genetics, lineage analysis and immunohistochemistry. The developmental role of C. elegans SEL-14 homologs will also be addressed by identification of deletion alleles, expression analysis, characterization of mutant phenotypes, and genetic analysis. The second aim tests the hypothesis that SEL-14 directly interacts with Notch receptors and cooperates with classical DSL ligands in receptor activation. This model suggested by in vivo studies and will be tested in vitro using two-hybrid studies and tissue culture experiments. We will also examine the role of a previously uncharacterized, conserved sequence motif that is found only in SEL-14, C. elegans homologs and classical Notch ligands. Structure/function studies will address the function of this conserved motif in SEL-14 and in classical Notch ligands. Understanding Notch ligands is critical given the essential role of Notch signaling in development, stem cell maintenance, cancer and memory. Defects in Notch signaling and presenilin processing cause numerous diseases. Deltas has been implicated in spondylocostal dysostosis. Mutations in NotchS and Jaggedl cause CADASIL and Alagille syndromes, respectively. These are dominantly inherited disorders associated with stroke and dementia. Notch and APR not only share common processing pathways like presenilins, but recent studies suggest that Notch and APR may both contribute to Alzheimer's disease pathology like presenilins. All of these disorders are poorly understood and no effective treatment is available. We anticipate that understanding ligand regulation of Notch signaling will be a first step in the development of therapies for these and other disorders.

描述（由申请人提供）：保守的Notch信号通路对于脊椎动物和无脊椎动物发育期间的细胞命运特化和侧向抑制至关重要。Notch受体结合细胞外配体（例如Delta、Serrate和LAG-2），所述配体含有对于受体活化至关重要的保守DSL结构域。我们在C.在脊椎动物中具有同源物的秀丽线虫。与典型的Notch配体不同，这些蛋白质缺乏保守的DSL结构域，但它们与经典的Notch配体共享保守的序列基序。本提案的第一个目的是详细研究这些新的推定配体之一的功能。SEL-14是正常发育所必需的，初步研究表明它激活Notch信号传导。SEL-14在Notch信号传导中的作用将使用遗传学、谱系分析和免疫组织化学在分子和细胞水平上进行检查。C.也将通过缺失等位基因的鉴定、表达分析、突变体表型的表征和遗传分析来解决线虫SEL-14同源物。第二个目的测试SEL-14直接与Notch受体相互作用并与经典DSL配体在受体活化中合作的假设。该模型由体内研究提出，并将使用双杂交研究和组织培养实验在体外进行测试。我们还将研究一个以前未表征的保守序列基序的作用，该基序仅在SEL-14，C。elegans同源物和经典Notch配体。结构/功能研究将解决SEL-14和经典Notch配体中该保守基序的功能。了解Notch配体至关重要，因为Notch信号在发育，干细胞维持，癌症和记忆中起着重要作用。Notch信号传导和早老素加工的缺陷导致许多疾病。三角洲与脊椎肋骨发育不全有关。NotchS和Jaggedl的突变分别导致CADASIL和Alagille综合征。这些是与中风和痴呆相关的显性遗传性疾病。Notch和APR不仅像早老素一样共享共同的加工途径，而且最近的研究表明，Notch和APR可能都像早老素一样有助于阿尔茨海默病的病理学。所有这些疾病都知之甚少，没有有效的治疗方法。我们预计，了解Notch信号的配体调节将是开发这些和其他疾病治疗方法的第一步。