果蝇微管互作蛋白的大规模筛选研究

Large-scale screening of microtubule interacting-proteins in Drosophila

微管是一种具有极性的、管状的细胞内动态结构,是细胞骨架的重要组分.一些跟微管相关的基因发生突变时,有可能致使严重的人类疾病的发生.这些相关基因编码的蛋白即为微管互作蛋白.影响微管的基因众多,目前发现的影响微管正常组装的蛋白还只是冰山一角,仍有诸多影响微管的互作蛋白等待人类“挖掘”.我们利用已有的果蝇RNAi文库,通过UAS/Gal4系统,对部分基因进行敲减(RNAi),使基因在果蝇幼虫肌肉中特异性沉默,经过免疫染色后观察这些基因功能敲降果蝇的肌肉微管形态,以此来筛选微管互作蛋白.我们共鉴定了541个基因,筛选出微管有表型的40个.其中一些基因在线粒体、内质网、高尔基体、过氧化物酶体等细胞器中具有特定的功能.因此,我们的筛选工作不仅为构建微管相关疾病模型提供了铺垫,为微管相关疾病治疗工作提供了一定的帮助;而且对细胞结构中微管的非中心体微管组织中心探究提供了思路.

Microtubules are polar tubules,which are important components of cytoskeleton.Mutations in some genes associated with microtubules have the potential to cause serious human disease.The proteins encoded by these genes are called microtubule interaction proteins.There are many genes that affect microtubules.The proteins that affect the normal assembly of microtubules are only the tip of the iceberg,and there are still many interacting proteins that affect microtubules waiting to be explored.We used the existing Drosophila RNAi library to conduct partial gene knockdown(RNAi)through UAS/Gal4 system to make genes specifically silenced in drosophila larva muscle.After immunostaining,the muscle microtubule morphology of these gene function knockdown drosophilas were observed to screen the microtubule interaction proteins.A total of 541 genes were identified,and 40 genes were screened for microtubule phenotype.Some of these genes had specific functions in mitochondria,endoplasmic reticulum,golgi apparatus,peroxisome and other organelles.Therefore,our screening work not only provides a foundation for the construction of microtubule-related disease model,but also provides some help for the treatment of microtubule-related disease.It also provides ideas for exploring the non-centrosomal microtubule organization center in cell structure.