Ubiquitination at the lysine 27 residue of the Parkin ubiquitin-like domain is suggestive of a new mechanism of Parkin activation.
Ubiquitination at the lysine 27 residue of the Parkin ubiquitin-like domain is suggestive of a new mechanism of Parkin activation.
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DOI：
10.1093/hmg/ddac064
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发表时间：
2022-08-17
影响因子：
3.5
通讯作者：
中科院分区：
文献类型：
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线粒体激酶PTEN诱导的激酶1(PINK1)和胞浆泛素连接酶(E3)Parkin/PRKN参与了线粒体的质量控制反应。PINK1在丝氨酸65处磷酸化泛素和Parkin泛素样(Ubl)结构域，促进Parkin激活和易位到受损线粒体。在Parkin激活后，Ub1结构域在赖氨酸(K)27和K48残基泛素化。然而，K27/K48泛素化对Parkin活性的贡献仍不清楚。在本研究中，通过产生果蝇Parkin(DParkin)突变体来阻断K56(对应于人类的K27)、K77(人类的K48)或两者的泛素化，以检测Parkin Ubl结构域泛素化对果蝇Parkin激活的影响。用精氨酸取代K56的dParkin(DParkin K56R)通过与PINK1的共表达挽救了果蝇的死亡，而dParkin K77R不能。DParkin K56R表现出线粒体断裂和运动停滞的能力降低，这分别是通过降解Parkin E3底物Mitofusin和Miro来实现的。与dParkin K56R不同，致病的dParkin K56N破坏了该蛋白的稳定性，这表明dParkin K56N不仅在K56位被非泛素修饰，而且激活的Ub1结构域的结构也受到了很大的影响。在PINK1介导的Parkin激活过程中，连接到Ub1结构域K27上的泛素可能被磷酸化，因为人Parkin K27R削弱了Parkin自身结合和反式激活。因此，我们的发现提示了一种新的Parkin激活机制，即通过磷酸化泛素连接到Parkin Ub1结构域的K27残基上形成激活复合体。
The mitochondrial kinase PTEN-induced kinase 1 (PINK1) and cytosolic ubiquitin ligase (E3) Parkin/PRKN are involved in mitochondrial quality control responses. PINK1 phosphorylates ubiquitin and the Parkin ubiquitin-like (Ubl) domain at serine 65 and promotes Parkin activation and translocation to damaged mitochondria. Upon Parkin activation, the Ubl domain is ubiquitinated at lysine (K) 27 and K48 residues. However, the contribution of K27/K48 ubiquitination toward Parkin activity remains unclear. In this study, ubiquitination of K56 (corresponding to K27 in the human), K77 (K48 in the human) or both was blocked by generating Drosophila Parkin (dParkin) mutants to examine the effects of Parkin Ubl domain ubiquitination on Parkin activation in Drosophila. The dParkin, in which K56 was replaced with arginine (dParkin K56R), rescued pupal lethality in flies by co-expression with PINK1, whereas dParkin K77R could not. The dParkin K56R exhibited reduced abilities of mitochondrial fragmentation and motility arrest, which are mediated by degrading Parkin E3 substrates Mitofusin and Miro, respectively. Pathogenic dParkin K56N, unlike dParkin K56R, destabilized the protein, suggesting that not only was dParkin K56N non-ubiquitin-modified at K56, but also the structure of the Ubl domain for activation was largely affected. Ubiquitin attached to K27 of the Ubl domain during PINK1-mediated Parkin activation was likely to be phosphorylated because human Parkin K27R weakened Parkin self-binding and activation in trans. Therefore, our findings suggest a new mechanism of Parkin activation, where an activation complex is formed through phospho-ubiquitin attachment on the K27 residue of the Parkin Ubl domain.
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影响因子：
4.5
作者：
Liu S;Sawada T;Lee S;Yu W;Silverio G;Alapatt P;Millan I;Shen A;Saxton W;Kanao T;Takahashi R;Hattori N;Imai Y;Lu B
通讯作者：
Lu B
影响因子：
7.7
作者：
Kazlauskaite A;Martínez-Torres RJ;Wilkie S;Kumar A;Peltier J;Gonzalez A;Johnson C;Zhang J;Hope AG;Peggie M;Trost M;van Aalten DM;Alessi DR;Prescott AR;Knebel A;Walden H;Muqit MM
通讯作者：
Muqit MM
影响因子：
5.8
作者：
Kazlauskaite A;Kelly V;Johnson C;Baillie C;Hastie CJ;Peggie M;Macartney T;Woodroof HI;Alessi DR;Pedrioli PG;Muqit MM
通讯作者：
Muqit MM
影响因子：
64.8
作者：
Gladkova C;Maslen SL;Skehel JM;Komander D
通讯作者：
Komander D
影响因子：
14.9
作者：
Moriya Y;Kawano S;Okuda S;Watanabe Y;Matsumoto M;Takami T;Kobayashi D;Yamanouchi Y;Araki N;Yoshizawa AC;Tabata T;Iwasaki M;Sugiyama N;Tanaka S;Goto S;Ishihama Y
通讯作者：
Ishihama Y