smn-1与uaf-1相互作用调节秀丽隐杆线虫寿命和运动功能
发布时间：2018-07-24 15:33
【摘要】：脊髓性肌萎缩症(Spinal Muscular Atrophy, SMA)是最常见的致死性神经肌肉疾病,由运动神经元生存基因SMN1(Survival Motor Neuron1)的功能缺失型突变引起,其具体机制尚不清楚。有研究表明,SMN1影响U snRNP (small nuclear RiboNucleoProtein)的组装,U snRNP是执行RNA剪接的主要分子复合物,RNA剪接缺陷可能是造成SMA的原因。 目的： 研究SMNl的秀丽隐杆线虫同源基因smmn-1在RNA剪接中的功能以及smmn-1与识别3’剪接位点的重要剪接因子uaf-1的相互作用。 方法： 1.tos-1是一个敏感的线虫内源性剪接报告基因。我们利用RT-PCR检测了smn-1的无效突变smn-1(ok355△)对tos-1剪接的影响,并通过RNAi减少smmn-1的表达,确认tos-1的剪接变化。同时,利用tos-13’剪接位点突变质粒注射产生转基因的smn-1(ok355△)线虫,研究强3’剪接位点的识别是否需要smn-1。 2.smn-1(ok355△)线虫发育停滞在幼虫晚期,寿命缩短,并伴有运动缺陷。为了分析smmn-1与uaf-1的相互作用,我们检测了相关线虫的寿命和三个运动指标,包括bodybends、thrashing和咽部pumping。 3.U snRNA是U snRNP的核心组分,参与RNA剪接的U snRNA有五种,分别为U1、U2、U4、U5和U6snRNA。因此我们利用RT-qPCR检测了smn-1(ok355A)突变体中U snRNA的表达。 结果： 1.smn-1(ok355△)突变改变tos-1的剪接,tos-1中弱3’剪接位点的识别需要smn-1,弱3’剪接位点的识别不需要smn-1。 2.uaf-1突变延长smn-1(ok355△)突变体的寿命并改善其运动功能。 3.在smn-1(ok355△)突变体中,我们检测到U1和U5snRNA的表达下降,U2、U4和U6snRNA的表达上升。 结论： 在线虫中,RNA的正常剪接需要smn-1, smn-1是一个重要的剪接因子；uaf-1是smn-1的修饰基因,两者相互作用调节线虫寿命和运动功能；smn-1(ok355△)突变影响参与RNA剪接的重要分子UsnRNA的表达。
[Abstract]:Spinal muscular atrophy (Spinal Muscular Atrophy, SMA) is the most common fatal neuromuscular disease caused by the deletion of motor neuron survival gene (SMN1 (Survival Motor Neuron1). Some studies have shown that the RNA splicing defects of SMMN1, a major molecular complex that performs RNA splicing, may be the cause of SMA. Aim: to study the function of SMNl homologous gene smmn-1 in RNA splicing and the interaction between smmn-1 and uaf-1, an important splicing factor that recognizes 3 'splicing sites. Methods: 1.tos-1 is a sensitive endogenous splicing reporter gene of nematodes. We use RT-PCR to detect the effect of smn-1 (ok355) on tos-1 splicing and confirm the change of tos-1 splicing by reducing the expression of smmn-1 by RNAi. At the same time, the mutant plasmid of tos-13 'splicing site was injected to produce transgenic smn-1 (ok355) nematode. Whether the identification of strong 3'splicing sites required smn-1. 2.smn-1 (ok355) nematode development was stopped in the late larval stage, and the life span of 2.smn-1 (ok355) nematode was shortened. Accompanied by motion defects. In order to analyze the interaction between smmn-1 and uaf-1, we examined the longevity and three movement indexes of nematodes, including body bendsthrashing and pharyngeal pumping.3.U snRNA is the core component of U snRNP. There are five kinds of U snRNA involved in RNA splicing, which are U1U2U2U4U5 and U6snRNA. Therefore, we used RT-qPCR to detect the expression of U snRNA in smn-1 (ok355A) mutants. Results: 1.smn-1 (ok355) mutation required smn-1 to recognize weak 3 'splicing sites in tos-1 splicing tos-1, while smn-1 was not required for weak 3' splicing sites. 2.uaf-1 mutation prolonged the life span of smn-1 (ok355) mutants and improved them. Motor function. In smn-1 (ok355) mutants, we found that the expression of U1 and U5snRNA decreased and the expression of U2U4 and U6snRNA increased. Conclusion: Smn-1 is required for the normal splicing of smn-1 in online worms. Smn-1 is an important splicing factor, which is a modified gene of smn-1. The interaction between them regulates the longevity and motor function of nematodes. Smn-1 (ok355) mutation affects the expression of UsnRNA, an important molecule involved in RNA splicing.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R746.4
本文编号：2141832
[Abstract]:Spinal muscular atrophy (Spinal Muscular Atrophy, SMA) is the most common fatal neuromuscular disease caused by the deletion of motor neuron survival gene (SMN1 (Survival Motor Neuron1). Some studies have shown that the RNA splicing defects of SMMN1, a major molecular complex that performs RNA splicing, may be the cause of SMA. Aim: to study the function of SMNl homologous gene smmn-1 in RNA splicing and the interaction between smmn-1 and uaf-1, an important splicing factor that recognizes 3 'splicing sites. Methods: 1.tos-1 is a sensitive endogenous splicing reporter gene of nematodes. We use RT-PCR to detect the effect of smn-1 (ok355) on tos-1 splicing and confirm the change of tos-1 splicing by reducing the expression of smmn-1 by RNAi. At the same time, the mutant plasmid of tos-13 'splicing site was injected to produce transgenic smn-1 (ok355) nematode. Whether the identification of strong 3'splicing sites required smn-1. 2.smn-1 (ok355) nematode development was stopped in the late larval stage, and the life span of 2.smn-1 (ok355) nematode was shortened. Accompanied by motion defects. In order to analyze the interaction between smmn-1 and uaf-1, we examined the longevity and three movement indexes of nematodes, including body bendsthrashing and pharyngeal pumping.3.U snRNA is the core component of U snRNP. There are five kinds of U snRNA involved in RNA splicing, which are U1U2U2U4U5 and U6snRNA. Therefore, we used RT-qPCR to detect the expression of U snRNA in smn-1 (ok355A) mutants. Results: 1.smn-1 (ok355) mutation required smn-1 to recognize weak 3 'splicing sites in tos-1 splicing tos-1, while smn-1 was not required for weak 3' splicing sites. 2.uaf-1 mutation prolonged the life span of smn-1 (ok355) mutants and improved them. Motor function. In smn-1 (ok355) mutants, we found that the expression of U1 and U5snRNA decreased and the expression of U2U4 and U6snRNA increased. Conclusion: Smn-1 is required for the normal splicing of smn-1 in online worms. Smn-1 is an important splicing factor, which is a modified gene of smn-1. The interaction between them regulates the longevity and motor function of nematodes. Smn-1 (ok355) mutation affects the expression of UsnRNA, an important molecule involved in RNA splicing.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R746.4
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