Cavadini P, Gellera C, Patel P I, Isaya G
Department of Pediatric and Adolescent Medicine, Mayo Clinic and Foundation, 200 First Street SW, Rochester, MN 55905, USA.
Hum Mol Genet. 2000 Oct 12;9(17):2523-30. doi: 10.1093/hmg/9.17.2523.
Frataxin is a nuclear-encoded mitochondrial protein widely conserved among eukaryotes. Human frataxin (fxn) is severely reduced in Friedreich ataxia (FRDA), a frequent autosomal recessive neuro- and cardio-degenerative disease. Whereas the function of fxn is unknown, the yeast frataxin homolog (Yfh1p) has been shown to be involved in mitochondrial iron homeostasis and protection from free radical toxicity. Evidence of iron accumulation and oxidative damage in cardiac tissue from FRDA patients suggests that fxn may have a similar function, but whether yeast and human frataxin actually have interchangeable roles in mitochondrial iron homeostasis is unknown. We show that a wild-type FRDA cDNA can complement Yfh1p-deficient yeast (yfh1 delta) by preventing the mitochondrial iron accumulation and oxidative damage associated with loss of Yfh1p. We analyze the functional effects of two FRDA point mutations, G130V and W173G, associated with a mild and a severe clinical presentation, respectively. The G130V mutation affects protein stability and results in low levels of mature (m) fxn, which are nevertheless sufficient to rescue yfh1 delta yeast. The W173G mutation affects protein processing and stability and results in severe m-fxn deficiency. Expression of the FRDA (W173G) cDNA in yfh1 delta yeast leads to increased levels of mitochondrial iron which are not as elevated as in Yfh1p-deficient cells but are above the threshold for oxidative damage of mitochondrial DNA and iron-sulfur centers, causing a typical yfh1 delta phenotype. These results demonstrate that fxn functions like Yfh1p, providing experimental support to the hypothesis that FRDA is a disorder of mitochondrial iron homeostasis.
铁调素是一种核编码的线粒体蛋白,在真核生物中广泛保守。人类铁调素(fxn)在弗里德赖希共济失调(FRDA)中严重减少,FRDA是一种常见的常染色体隐性神经和心脏退行性疾病。虽然fxn的功能尚不清楚,但酵母铁调素同源物(Yfh1p)已被证明参与线粒体铁稳态和免受自由基毒性的影响。FRDA患者心脏组织中铁积累和氧化损伤的证据表明fxn可能具有类似的功能,但酵母和人类铁调素在线粒体铁稳态中是否真的具有可互换的作用尚不清楚。我们表明,野生型FRDA cDNA可以通过防止与Yfh1p缺失相关的线粒体铁积累和氧化损伤来补充Yfh1p缺陷的酵母(yfh1 delta)。我们分析了两个与轻度和重度临床表现相关的FRDA点突变G130V和W173G的功能影响。G130V突变影响蛋白质稳定性,导致成熟(m)fxn水平较低,但仍足以拯救yfh1 delta酵母。W173G突变影响蛋白质加工和稳定性,导致严重的m-fxn缺陷。yfh1 delta酵母中FRDA(W173G)cDNA的表达导致线粒体铁水平升高,虽然不如Yfh1p缺陷细胞中升高得那么高,但高于线粒体DNA和铁硫中心氧化损伤的阈值,导致典型的yfh1 delta表型。这些结果表明fxn的功能与Yfh1p相似,为FRDA是线粒体铁稳态紊乱这一假设提供了实验支持。