Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI, USA.
Department of Neurology, University of Michigan Medical School, Ann Arbor, MI, USA.
Neurobiol Dis. 2019 Dec;132:104535. doi: 10.1016/j.nbd.2019.104535. Epub 2019 Jul 13.
The most commonly inherited dominant ataxia, Spinocerebellar Ataxia Type 3 (SCA3), is caused by a CAG repeat expansion that encodes an abnormally long polyglutamine (polyQ) repeat in the disease protein ataxin-3, a deubiquitinase. Two major full-length isoforms of ataxin-3 exist, both of which contain the same N-terminal portion and polyQ repeat, but differ in their C-termini; one (denoted here as isoform 1) contains a motif that binds ataxin-3's substrate, ubiquitin, whereas the other (denoted here as isoform 2) has a hydrophobic tail. Most SCA3 studies have focused on isoform 1, the predominant version in mammalian brain, yet both isoforms are present in brain and a better understanding of their relative pathogenicity in vivo is needed. We took advantage of the fruit fly, Drosophila melanogaster to model SCA3 and to examine the toxicity of each ataxin-3 isoform. Our assays reveal isoform 1 to be markedly more toxic than isoform 2 in all fly tissues. Reduced toxicity from isoform 2 is due to much lower protein levels as a result of its expedited degradation. Additional studies indicate that isoform 1 is more aggregation-prone than isoform 2 and that the C-terminus of isoform 2 is critical for its enhanced proteasomal degradation. According to our results, although both full-length, pathogenic ataxin-3 isoforms are toxic, isoform 1 is likely the primary contributor to SCA3 due to its presence at higher levels. Isoform 2, as a result of rapid degradation that is dictated by its tail, is unlikely to be a key player in this disease. Our findings provide new insight into the biology of this ataxia and the cellular processing of the underlying disease protein.
最常见的显性遗传性共济失调,脊髓小脑性共济失调 3 型(SCA3),是由 CAG 重复扩展引起的,该扩展在疾病蛋白 ataxin-3 中编码异常长的多聚谷氨酰胺(polyQ)重复,ataxin-3 是一种去泛素化酶。ataxin-3 存在两种主要的全长异构体,它们都包含相同的 N 端部分和 polyQ 重复,但在 C 端不同;一个(这里表示为异构体 1)包含一个结合 ataxin-3 底物泛素的基序,而另一个(这里表示为异构体 2)则具有疏水性尾巴。大多数 SCA3 研究都集中在哺乳动物大脑中主要存在的异构体 1 上,但两种异构体都存在于大脑中,需要更好地了解它们在体内的相对致病性。我们利用果蝇,黑腹果蝇来模拟 SCA3 并研究每个 ataxin-3 异构体的毒性。我们的实验表明,在所有的果蝇组织中,异构体 1 的毒性明显高于异构体 2。异构体 2 毒性降低是由于其快速降解导致的蛋白水平降低。进一步的研究表明,异构体 1 比异构体 2 更容易聚集,并且异构体 2 的 C 端对于其增强的蛋白酶体降解至关重要。根据我们的结果,尽管全长致病性 ataxin-3 异构体都具有毒性,但由于其存在水平较高,异构体 1 可能是 SCA3 的主要贡献者。由于其尾部决定的快速降解,异构体 2 不太可能成为该疾病的关键因素。我们的研究结果为该共济失调的生物学和潜在疾病蛋白的细胞处理提供了新的见解。