IDENTIFICATION OF NOVEL CHONDROITIN PROTEOGLYCANS IN CAENORHABDITIS ELEGANS

秀丽隐杆线虫中新型软骨素蛋白聚糖的鉴定

• 批准号: 7602151
• 负责人: Jeffrey D Esko
• 金额: $ 0.62万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602151
• 关键词: Binding; Biochemical; Caenorhabditis elegans; Chitin; Chondroitin; Chondroitin Sulfate Proteoglycan; Computer Retrieval of Information on Scientific Projects Database; Core Protein; Cytokinesis; Development; Drosophila melanogaster; Embryo; Embryonic Development; Funding; Grant; Homologous Gene; Hydra Polyps; Individual; Inorganic Sulfates; Institution; Invertebrates; Mass Spectrum Analysis; Mechanics; Morphogenesis; Nematoda; Phenocopy; Play; Proteoglycan; RNA Interference; Research; Research Personnel; Resources; Role; Source; Tissues; United States National Institutes of Health; Unspecified or Sulfate Ion Sulfates; Vertebrates; Western Blotting; chondroitin synthase; egg; embryo cell; novel; proteoglycan core protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Vertebrates produce multiple chondroitin sulfate proteoglycans that play important roles in development and tissue mechanics. In the nematode Caenorhabditis elegans, the chondroitin chains lack sulfate but nevertheless play essential roles in embryonic development and vulval morphogenesis. However, assignment of these functions to specific proteoglycans has been limited by the lack of identified core proteins. We used a combination of biochemical purification, Western blotting, and mass spectrometry to identify nine C. elegans chondroitin proteoglycan core proteins, none of which have homologues in vertebrates or other invertebrates such as Drosophila melanogaster or Hydra vulgaris. CPG-1/CEJ-1 and CPG-2 are expressed during embryonic development and bind chitin, suggesting a structural role in the egg. RNA interference (RNAi) depletion of individual CPGs had no effect on embryonic viability, but simultaneous depletion of CPG-1/CEJ-1 and CPG-2 resulted in multinucleated single-cell embryos. This embryonic lethality phenocopies RNAi depletion of the SQV-5 chondroitin synthase, suggesting that chondroitin chains on these two proteoglycans are required for cytokinesis.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 脊椎动物产生多种硫酸软骨素蛋白聚糖，在发育和组织力学中发挥重要作用。在秀丽隐杆线虫中，软骨素链缺乏硫酸盐，但在胚胎发育和外阴形态发生中发挥重要作用。然而，这些功能分配给特定的蛋白聚糖已被确定的核心蛋白的缺乏限制。我们采用生化纯化、蛋白质印迹和质谱分析相结合的方法鉴定了9种C。线虫软骨素蛋白聚糖核心蛋白，它们在脊椎动物或其他无脊椎动物如黑腹果蝇或水螅中没有同源物。CPG-1/CEJ-1和CPG-2在胚胎发育过程中表达，并结合几丁质，表明在卵中的结构作用。RNA干扰（RNAi）消耗单个CPG对胚胎存活率没有影响，但同时消耗CPG-1/CEJ-1和CPG-2导致多核单细胞胚胎。这种胚胎致死性表型模仿SQV-5软骨素合酶的RNAi耗竭，表明这两种蛋白聚糖上的软骨素链是胞质分裂所需的。