The complex I subunit NDUFA10 selectively rescues Drosophila pink1 mutants through a mechanism independent of mitophagy.
The complex I subunit NDUFA10 selectively rescues Drosophila pink1 mutants through a mechanism independent of mitophagy.
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DOI：
10.1371/journal.pgen.1004815
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发表时间：
2014-11
期刊：
影响因子：
4.5
通讯作者：
Whitworth AJ
中科院分区：
文献类型：
--
作者：
Pogson JH;Ivatt RM;Sanchez-Martinez A;Tufi R;Wilson E;Mortiboys H;Whitworth AJ
PINK 1是一种靶向丝氨酸/苏氨酸激酶，其突变可导致常染色体隐性遗传帕金森病（PD）。大量证据表明，PINK 1与另一个PD基因parkin一起调节线粒体形态和线粒体自噬。然而，PINK 1的缺失也导致复合物I（CI）缺陷，并且最近被认为通过NDUFA 10/ND 42亚基的磷酸化来调节CI。为了进一步探索PINK 1和Parkin影响线粒体完整性的机制，我们在果蝇细胞中进行了一项筛选，以寻找由pink 1 RNAi引起的表型复制或抑制线粒体过度融合的基因。从该筛选回收的基因中有ND 42。在果蝇pink 1突变体中，转基因过量表达ND 42或其共伴侣西西里足以恢复CI活性，并部分挽救几种表型，包括飞行和攀爬缺陷和飞行肌肉中的线粒体破坏。在此，CI活性的恢复和运动的部分拯救似乎对ND 42在Ser-250处的磷酸化没有特定要求。与pink 1突变体相反，ND 42或西西里的过表达未能拯救任何果蝇parkin突变体表型。我们还发现，敲低人类同源物NDUFA 10，仅最小程度地影响CCCP诱导的线粒体自噬，并且过表达NDUFA 10不能恢复PINK 1丢失后的Parkin细胞易位。这些结果表明，体内拯救是由于恢复CI活性，而不是促进线粒体自噬。我们的研究结果支持了一种新的观点，即PINK 1在调节CI活性中起作用，而不是在线粒体自噬中与Parkin一起起作用。与神经退行性运动障碍帕金森病（PD）的遗传形式相关的两个基因，PINK 1和parkin，通过包括功能失调的线粒体的降解（称为线粒体自噬）和复合物I（CI）活性的维持的机制在线粒体稳态中起重要作用。在这里，我们报告了在果蝇细胞中基于RNAi筛选可能调节PINK 1-Parkin通路的基因的结果，该基因鉴定了CI的亚基NDUFA 10（果蝇中的ND 42）。使用一个完善的细胞系统和体内果蝇遗传学，我们证明，而NDUFA 10/ND 42只发挥了最小的作用，在线粒体自噬，恢复CI活动通过过度表达ND 42或其co-chaperone西西里岛是能够大大拯救行为缺陷pink 1突变体，但不是parkin突变体。此外，虽然parkin过表达已知拯救pink 1突变体，但它显然在不恢复CI活性的情况下实现了这一点。这些结果表明，增加CI活性或促进线粒体自噬在pink 1突变体中可能是有益的，并进一步强调了PINK 1和Parkin的可分离功能。
Mutations in PINK1, a mitochondrially targeted serine/threonine kinase, cause autosomal recessive Parkinson's disease (PD). Substantial evidence indicates that PINK1 acts with another PD gene, parkin, to regulate mitochondrial morphology and mitophagy. However, loss of PINK1 also causes complex I (CI) deficiency, and has recently been suggested to regulate CI through phosphorylation of NDUFA10/ND42 subunit. To further explore the mechanisms by which PINK1 and Parkin influence mitochondrial integrity, we conducted a screen in Drosophila cells for genes that either phenocopy or suppress mitochondrial hyperfusion caused by pink1 RNAi. Among the genes recovered from this screen was ND42. In Drosophila pink1 mutants, transgenic overexpression of ND42 or its co-chaperone sicily was sufficient to restore CI activity and partially rescue several phenotypes including flight and climbing deficits and mitochondrial disruption in flight muscles. Here, the restoration of CI activity and partial rescue of locomotion does not appear to have a specific requirement for phosphorylation of ND42 at Ser-250. In contrast to pink1 mutants, overexpression of ND42 or sicily failed to rescue any Drosophila parkin mutant phenotypes. We also find that knockdown of the human homologue, NDUFA10, only minimally affecting CCCP-induced mitophagy, and overexpression of NDUFA10 fails to restore Parkin mitochondrial-translocation upon PINK1 loss. These results indicate that the in vivo rescue is due to restoring CI activity rather than promoting mitophagy. Our findings support the emerging view that PINK1 plays a role in regulating CI activity separate from its role with Parkin in mitophagy. Two genes linked to heritable forms of the neurodegenerative movement disorder Parkinson's disease (PD), PINK1 and parkin, play important roles in mitochondrial homeostasis through mechanisms which include the degradation of dysfunctional mitochondria, termed mitophagy, and the maintenance of complex I (CI) activity. Here we report the findings of an RNAi based screen in Drosophila cells for genes that may regulate the PINK1-Parkin pathway which identified NDUFA10 (ND42 in Drosophila), a subunit of CI. Using a well-established cellular system and in vivo Drosophila genetics, we demonstrate that while NDUFA10/ND42 only plays a minimal role in mitophagy, restoration of CI activity through overexpression of either ND42 or its co-chaperone sicily is able to substantially rescue behavioral deficits in pink1 mutants but not parkin mutants. Moreover, while parkin overexpression is known to rescue pink1 mutants, it apparently achieves this without restoring CI activity. These results suggest that increasing CI activity or promoting mitophagy can be beneficial in pink1 mutants, and further highlights separable functions of PINK1 and Parkin.
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DOI：
10.1073/pnas.1321207111
发表时间：
2014-06-10
影响因子：
11.1
作者：
Ivatt, Rachael M.;Sanchez-Martinez, Alvaro;Whitworth, Alexander J.
通讯作者：
Whitworth, Alexander J.
DOI：
10.1006/bmmb.1994.1004
发表时间：
1994-02-01
期刊：
BIOCHEMICAL MEDICINE AND METABOLIC BIOLOGY
影响因子：
--
作者：
BIRCHMACHIN, MA;BRIGGS, HL;TURNBULL, DM
通讯作者：
TURNBULL, DM
DOI：
10.1016/j.bbadis.2013.01.004
发表时间：
2013-04-01
影响因子：
6.2
作者：
Cali, Tito;Ottolini, Denis;Brini, Marisa
通讯作者：
Brini, Marisa
影响因子：
3.9
作者：
Krebs, Catharine E.;Karkheiran, Siamak;Powell, James C.;Cao, Mian;Makarov, Vladimir;Darvish, Hossein;Di Paolo, Gilbert;Walker, Ruth H.;Shahidi, Gholam Ali;Buxbaum, Joseph D.;De Camilli, Pietro;Yue, Zhenyu;Paisan-Ruiz, Coro
通讯作者：
Paisan-Ruiz, Coro
DOI：
10.1083/jcb.200910140
发表时间：
2010-04-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Matsuda N;Sato S;Shiba K;Okatsu K;Saisho K;Gautier CA;Sou YS;Saiki S;Kawajiri S;Sato F;Kimura M;Komatsu M;Hattori N;Tanaka K
通讯作者：
Tanaka K