Expanding Excellence in Developmental Biology in Oklahoma

中心粒重复（CD）基因的突变与原发性高血压等疾病的发病率相关， 小头畸形、原始侏儒症和癌症。然而，许多这些疾病相关的影响， CD基因突变对其各自蛋白功能的影响目前尚不清楚。该提案利用一个 保守CD基因纺锤体组装异常蛋白6中的这种疾病相关突变 （HsSAS-6/ SAS-6）作为研究SAS-6在纤毛发生和CD中的功能的工具。这种疾病相关的HsSAS- 6突变与原发性小头畸形的发生有关，对应于C. 优雅我们的长期目标是利用保守CD基因的疾病相关等位基因， 了解它们在调节纤毛发生和CD中的作用机制。这方面的总体目标 应用是确定sas-6（L 69 T）突变对C. elegans ciliogenesis和CD.的 中心假设是sas-6（L 69 T）突变抑制了SAS-6功能，导致纤毛发生受损 和CD。该项目的基本原理是，由于这种突变具有已知的病理后果， 它将为SAS-6的正常生物学功能提供重要的见解。中央 将通过追求两个特定目标来检验假设：1）确定sas-6（L 69 T）突变的影响 对秀丽隐阐明了sas-6（L 69 T）突变的分子机制， 在C中损害CD。优雅在第一个目标下，研究了sas-6（L 69 T）突变对C. elegans 纤毛发生将通过对C. 携带这种突变的线虫CRISPR菌株。在第二个目标中，各种细胞生物学和 将进行生化分析，以阐明这种突变影响的分子机制。 CD.在申请中提出的研究是创新的，因为i）这是第一项研究调查 原发性小头畸形相关SAS-6突变在多细胞真核动物模型中的作用。 ii）这是第一项使用疾病相关的sas-6等位基因来研究SAS-6在调节 纤毛发生为任何疾病制定治疗或预防干预措施， 了解疾病相关突变的基本后果， 科学这项拟议中的研究意义重大，因为目前还不知道 这种原发性小头畸形相关的SAS-6突变的机制作用。了解细胞和 sas-6（L 69 T）突变的生化后果是了解 原发性小头畸形的发病机制和进展。

Mutations in centriole duplication (CD) genes are correlated with the incidence of diseases such as primary microcephaly, primordial dwarfism and cancer. However, the effect of many of these disease-associated mutations in CD genes on their respective protein functions is currently unknown. This proposal utilizes one such disease-associated mutation in the conserved CD gene Spindle Assembly Abnormal Protein 6 (HsSAS-6/ SAS-6) as a tool to study SAS-6 function in ciliogenesis and CD. This disease-relevant HsSAS- 6 mutation is associated with the incidence of primary microcephaly and corresponds to sas-6(L69T) in C. elegans. Our long-term goal is to exploit disease-associated alleles of conserved CD genes to better understand their mechanism of action in regulating ciliogenesis and CD. The overall objectives in this application are to determine the effect of the sas-6(L69T) mutation on C. elegans ciliogenesis and CD. The central hypothesis is that the sas-6(L69T) mutation inhibits SAS-6 function leading to impaired ciliogenesis and CD. The rationale for this project is that since this mutation has a known pathological consequence in humans, it will provide important insights into the normal biological function of SAS-6. The central hypothesis will be tested by pursuing two specific aims: 1) Determine the effect of the sas-6(L69T) mutation on C. elegans ciliogenesis; 2) Elucidate the molecular mechanism by which the sas-6(L69T) mutation impairs CD in C. elegans. Under the first aim, the consequences of the sas-6(L69T) mutation on C. elegans ciliogenesis will be determined by performing a thorough microscopic and behavioral analysis of a C. elegans CRISPR strain carrying this mutation. In the second aim, a variety of cell biological and biochemical assays will be performed to clarify the molecular mechanism by which this mutation affects CD. The research proposed in the application is innovative because i) This is the first study investigating the effects of a primary-microcephaly-associated sas-6 mutation in a multicellular eukaryotic animal model. ii) This is the first study that uses a disease-associated sas-6 allele to study SAS-6 function in regulating ciliogenesis. Developing therapeutic or preventative interventions for any disease begins with understanding the fundamental consequences of disease-associated mutations at the level of basic science. The proposed research is significant because there is currently nothing known about the mechanistic effects of this primary microcephaly-associated sas-6 mutation. Understanding the cellular and biochemical consequences of the sas-6(L69T) mutation is the first important step to understanding the mechanism of incidence and progression of primary microcephaly.