Glycan alterations in C.elegans surface mutants

线虫表面突变体中的聚糖改变

• 批准号: 6660634
• 负责人: PATRICIA BERNINSONE
• 金额: $ 3.14万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-20 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6660634
• 关键词: Caenorhabditis elegans; N acetylglucosamine; biological models; cell cell interaction; cell membrane; double stranded RNA; gas chromatography mass spectrometry; gene expression; glycolipids; glycoprotein biosynthesis; glycoproteins; glycosylation; growth /development; lectin; matrix assisted laser desorption ionization; model design /development; molecular cloning; mutant; phenotype; pleiotropism; posttranslational modifications; proteoglycan; restriction fragment length polymorphism; single nucleotide polymorphism

DESCRIPTION (provided by applicant): Development of metazoans requires the production of multiple cell types and the morphogenesis of complex multicellular structures. Genetic studies have established roles for glycosylation and proteoglycan biosynthesis in developmental signaling. The nematode Caenorhabditis elegans is a good model system to study the role of glycoconjugates in multicellular organisms as well as a prototypic model for parasitic nematodes because of the knowledge of its complete genomic sequence and development. In this R2 I (Exploratory / Developmental) grant application, we wish to explore a novel potential functional linkage between expression of glycoconjugates and developmental events in C. elegans. Srf-9, srf-8 and srf-4 (surface) are single loci mutants that have multiple defects, including uncoordinated movement, protruding vulva, abnormal egg laying, and defective copulatory bursae and gonad morphology, yet they were isolated based on their ectopic surface binding to a lectin which binds to N-acetylglucosamine. Mutations in srf-9, srf-8 and srf-4 interact with mutations in the lin-12 gene, which a member of the LIN- 1 2/NOTCH family of receptor proteins that mediate cell-cell interactions to specify cell fate during development. We will test the hypothesis that the srf-9, srf-8 and srf-4 genes are involved in the biosynthesis, post-translational processing or secretion of glycoconjugates, and therefore link developmental events with glycoconjugates expression. Specifically, the proposed work will: 1) Define the glycoconjugates affected in srf-9, srf-8 and srf-4 mutants, through the structural characterization of cuticle glycan fractions, 2) Identify the srf-9, srf-8 and srf-4 genes through molecular cloning using a combination of Single Nucleotide Polymorphism (SNP) mapping, and marker rescue experiments. The information derived from these studies will provide the tools to study the role of these genes in developmental events in the nematode C. elegans. Because complex phenotypes are the hallmark of several human genetic disorders, including some that affect glycosylation, the proposed studies will establish the feasibility of using C. elegans srf mutants as a model for glycosylation disorders in humans.

描述（由申请人提供）：后生动物的发育需要多种细胞类型的产生和复杂多细胞结构的形态发生。遗传学研究已经确定了糖基化和蛋白聚糖生物合成在发育信号传导中的作用。线虫秀丽隐杆线虫是研究糖复合物在多细胞生物中的作用的良好模型系统，也是寄生线虫的原型模型，因为它了解其完整的基因组序列和发育。在此 R2 I（探索性/发展性）资助申请中，我们希望探索秀丽隐杆线虫糖缀合物表达与发育事件之间的新型潜在功能联系。 Srf-9、srf-8和srf-4（表面）是单基因座突变体，具有多种缺陷，包括运动不协调、外阴突出、产卵异常以及交配囊和性腺形态缺陷，但它们是根据其异位表面与凝集素结合而分离的，而凝集素与N-乙酰氨基葡萄糖结合。 srf-9、srf-8 和 srf-4 的突变与 lin-12 基因的突变相互作用，lin-12 基因是 LIN-1 2/NOTCH 受体蛋白家族的成员，可介导细胞与细胞之间的相互作用，从而在发育过程中指定细胞的命运。我们将检验以下假设：srf-9、srf-8 和 srf-4 基因参与糖复合物的生物合成、翻译后加工或分泌，因此将发育事件与糖复合物表达联系起来。具体来说，拟议的工作将：1) 通过角质层聚糖组分的结构表征，定义 srf-9、srf-8 和 srf-4 突变体中受影响的糖缀合物，2) 使用单核苷酸多态性 (SNP) 作图和标记拯救相结合的分子克隆，鉴定 srf-9、srf-8 和 srf-4 基因 实验。从这些研究中获得的信息将为研究这些基因在线虫发育事件中的作用提供工具。由于复杂的表型是几种人类遗传性疾病的标志，包括一些影响糖基化的疾病，因此拟议的研究将确定使用线虫 srf 突变体作为人类糖基化疾病模型的可行性。