Huynh Julie M, Dang Hope, Munoz-Tucker Isabel A, O'Ketch Marvin, Liu Ian T, Perno Savannah, Bhuyan Natasha, Crain Allison, Borbon Ivan, Fares Hanna
Department of Molecular and Cellular Biology, University of Arizona, Tucson, Arizona 85721.
Sahuaro High School, Tucson, Arizona 85710.
Genetics. 2016 Feb;202(2):619-38. doi: 10.1534/genetics.115.182485. Epub 2015 Nov 23.
Mutations in MCOLN1, which encodes the cation channel protein TRPML1, result in the neurodegenerative lysosomal storage disorder Mucolipidosis type IV. Mucolipidosis type IV patients show lysosomal dysfunction in many tissues and neuronal cell death. The ortholog of TRPML1 in Caenorhabditis elegans is CUP-5; loss of CUP-5 results in lysosomal dysfunction in many tissues and death of developing intestinal cells that results in embryonic lethality. We previously showed that a null mutation in the ATP-Binding Cassette transporter MRP-4 rescues the lysosomal defect and embryonic lethality of cup-5(null) worms. Here we show that reducing levels of the Endosomal Sorting Complex Required for Transport (ESCRT)-associated proteins DID-2, USP-50, and ALX-1/EGO-2, which mediate the final de-ubiquitination step of integral membrane proteins being sequestered into late endosomes, also almost fully suppresses cup-5(null) mutant lysosomal defects and embryonic lethality. Indeed, we show that MRP-4 protein is hypo-ubiquitinated in the absence of CUP-5 and that reducing levels of ESCRT-associated proteins suppresses this hypo-ubiquitination. Thus, increased ESCRT-associated de-ubiquitinating activity mediates the lysosomal defects and corresponding cell death phenotypes in the absence of CUP-5.
编码阳离子通道蛋白TRPML1的MCOLN1发生突变,会导致神经退行性溶酶体贮积症IV型。IV型粘脂贮积症患者在许多组织中表现出溶酶体功能障碍和神经元细胞死亡。秀丽隐杆线虫中TRPML1的直系同源物是CUP-5;CUP-5缺失会导致许多组织中的溶酶体功能障碍以及发育中的肠道细胞死亡,进而导致胚胎致死。我们之前表明,ATP结合盒转运蛋白MRP-4的无效突变可挽救cup-5(缺失)蠕虫的溶酶体缺陷和胚胎致死性。在此我们表明,降低参与转运所需的内体分选复合物(ESCRT)相关蛋白DID-2、USP-50和ALX-1/EGO-2的水平,也几乎能完全抑制cup-5(缺失)突变体的溶酶体缺陷和胚胎致死性,这些蛋白介导整合膜蛋白被隔离到晚期内体中的最终去泛素化步骤。实际上,我们表明在没有CUP-5的情况下MRP-4蛋白发生低泛素化,并且降低ESCRT相关蛋白的水平可抑制这种低泛素化。因此,在没有CUP-5的情况下,增加的ESCRT相关去泛素化活性介导了溶酶体缺陷和相应的细胞死亡表型。