Simonsen Anne, Cumming Robert C, Lindmo Karine, Galaviz Vanessa, Cheng Susan, Rusten Tor Erik, Finley Kim D
Department of Biochemistry, Center for Cancer Biomedicine, The Norwegian Radium Hospital, Montebello, 0310 Oslo, Norway.
Genetics. 2007 Jun;176(2):1283-97. doi: 10.1534/genetics.106.065011. Epub 2007 Apr 15.
Defects in lysosomal trafficking pathways lead to decreased cell viability and are associated with progressive disorders in humans. Previously we have found that loss-of-function (LOF) mutations in the Drosophila gene blue cheese (bchs) lead to reduced adult life span, increased neuronal death, and widespread CNS degeneration that is associated with the formation of ubiquitinated-protein aggregates. To identify potential genes that participate in the bchs functional pathway, we conducted a genetic modifier screen based on alterations of an eye phenotype that arises from high-level overexpression of Bchs. We found that mutations in select autophagic and endocytic trafficking genes, defects in cytoskeletal and motor proteins, as well as mutations in the SUMO and ubiquitin signaling pathways behave as modifiers of the Bchs gain-of-function (GOF) eye phenotype. Individual mutant alleles that produced viable adults were further examined for bchs-like phenotypes. Mutations in several lysosomal trafficking genes resulted in significantly decreased adult life spans and several mutants showed changes in ubiquitinated protein profiles as young adults. This work represents a novel approach to examine the role that lysosomal transport and function have on adult viability. The genes characterized in this study have direct human homologs, suggesting that similar defects in lysosomal transport may play a role in human health and age-related processes.
溶酶体运输途径的缺陷会导致细胞活力下降,并与人类的进行性疾病相关。此前我们发现,果蝇基因蓝纹奶酪(bchs)的功能丧失(LOF)突变会导致成虫寿命缩短、神经元死亡增加以及广泛的中枢神经系统退化,这与泛素化蛋白聚集体的形成有关。为了鉴定参与bchs功能途径的潜在基因,我们基于Bchs高水平过表达引起的眼睛表型改变进行了遗传修饰筛选。我们发现,选择的自噬和内吞运输基因中的突变、细胞骨架和运动蛋白的缺陷以及SUMO和泛素信号通路中的突变表现为Bchs功能获得(GOF)眼睛表型的修饰因子。对产生存活成虫的单个突变等位基因进一步检查其是否具有类似bchs的表型。几个溶酶体运输基因中的突变导致成虫寿命显著缩短,并且几个突变体在年轻成虫时泛素化蛋白谱发生了变化。这项工作代表了一种新的方法来研究溶酶体运输和功能对成虫活力的作用。本研究中鉴定的基因有直接的人类同源物,这表明溶酶体运输中的类似缺陷可能在人类健康和与年龄相关的过程中起作用。