An ALS-associated variant of the autophagy receptor SQSTM1/p62 reprograms binding selectivity toward the autophagy-related hATG8 proteins.
An ALS-associated variant of the autophagy receptor SQSTM1/p62 reprograms binding selectivity toward the autophagy-related hATG8 proteins.
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DOI：
10.1016/j.jbc.2021.101514
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Searle MS
中科院分区：
文献类型：
--
作者：
Brennan A;Layfield R;Long J;Williams HEL;Oldham NJ;Scott D;Searle MS
自噬受体对人自噬相关蛋白8（hATG 8）的识别是细胞质量控制系统中的关键步骤。自噬损伤是运动神经元疾病肌萎缩侧索硬化症（ALS）的一种致病机制。属于LC 3和GABARAP亚家族的hATG 8蛋白的重叠但特定的作用尚未完全理解，并且结合选择性通常被忽视。我们先前表明，ALS相关的SQSTM 1/p62（p62）自噬受体的变体在其ATG 8相互作用基序（AIM）中携带L341 V突变，在体外损害LC 3B的识别，产生自噬缺陷表型。对AIM识别hATG 8的理解的改进现在区分了LC 3相互作用和GABARAP相互作用基序，并预测L341 V取代的影响可能超出功能丧失，以偏置AIM结合偏好。通过生物物理分析，我们证实L341 V-AIM突变体与LC 3A、LC 3B、GABARAP和GABARAPL 1的结合受损。相反，p62 AIM与LC 3C和GABARAPL 2的相互作用不受该突变的影响。等温滴定量热法和NMR研究提供了对熵驱动的GABARAPL 2/p62相互作用以及L341 V突变如何被耐受的见解。竞争结合证明L341 V-AIM与一个hATG 8的结合减少，表现为与替代hATG 8的结合相对增加，表明hATG 8选择性的有效重编程。这些数据突出了单个AIM肽如何与不同的hATG 8竞争结合，并表明L341 V-AIM突变可能是新变体，代表可能延伸到其他人类疾病的疾病机制。
Recognition of human autophagy-related 8 (hATG8) proteins by autophagy receptors represents a critical step within this cellular quality control system. Autophagy impairment is known to be a pathogenic mechanism in the motor neuron disorder amyotrophic lateral sclerosis (ALS). Overlapping but specific roles of hATG8 proteins belonging to the LC3 and GABARAP subfamilies are incompletely understood, and binding selectivity is typically overlooked. We previously showed that an ALS-associated variant of the SQSTM1/p62 (p62) autophagy receptor bearing an L341V mutation within its ATG8-interacting motif (AIM) impairs recognition of LC3B in vitro, yielding an autophagy-deficient phenotype. Improvements in understanding of hATG8 recognition by AIMs now distinguish LC3-interaction and GABARAP-interaction motifs and predict the effects of L341V substitution may extend beyond loss of function to biasing AIM binding preference. Through biophysical analyses, we confirm impaired binding of the L341V-AIM mutant to LC3A, LC3B, GABARAP, and GABARAPL1. In contrast, p62 AIM interactions with LC3C and GABARAPL2 are unaffected by this mutation. Isothermal titration calorimetry and NMR investigations provided insights into the entropy-driven GABARAPL2/p62 interaction and how the L341V mutation may be tolerated. Competition binding demonstrated reduced association of the L341V-AIM with one hATG8 manifests as a relative increase in association with alternate hATG8s, indicating effective reprogramming of hATG8 selectivity. These data highlight how a single AIM peptide might compete for binding with different hATG8s and suggest that the L341V-AIM mutation may be neomorphic, representative of a disease mechanism that likely extends into other human disorders.
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DOI：
10.1126/science.aaa3650
发表时间：
2015-03-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Cirulli ET;Lasseigne BN;Petrovski S;Sapp PC;Dion PA;Leblond CS;Couthouis J;Lu YF;Wang Q;Krueger BJ;Ren Z;Keebler J;Han Y;Levy SE;Boone BE;Wimbish JR;Waite LL;Jones AL;Carulli JP;Day-Williams AG;Staropoli JF;Xin WW;Chesi A;Raphael AR;McKenna-Yasek D;Cady J;Vianney de Jong JM;Kenna KP;Smith BN;Topp S;Miller J;Gkazi A;FALS Sequencing Consortium;Al-Chalabi A;van den Berg LH;Veldink J;Silani V;Ticozzi N;Shaw CE;Baloh RH;Appel S;Simpson E;Lagier-Tourenne C;Pulst SM;Gibson S;Trojanowski JQ;Elman L;McCluskey L;Grossman M;Shneider NA;Chung WK;Ravits JM;Glass JD;Sims KB;Van Deerlin VM;Maniatis T;Hayes SD;Ordureau A;Swarup S;Landers J;Baas F;Allen AS;Bedlack RS;Harper JW;Gitler AD;Rouleau GA;Brown R;Harms MB;Cooper GM;Harris T;Myers RM;Goldstein DB
通讯作者：
Goldstein DB
影响因子：
3.5
作者：
Majcher, Veronika;Goode, Alice;Layfield, Robert
通讯作者：
Layfield, Robert
影响因子：
2.2
作者：
MONTELIONE, GT;WAGNER, G
通讯作者：
WAGNER, G
DOI：
10.1006/meth.1999.0852
发表时间：
1999-10-01
期刊：
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
影响因子：
--
作者：
Pierce, MM;Raman, CS;Nall, BT
通讯作者：
Nall, BT
影响因子：
19
作者：
Feng, Yuchen;Yao, Zhiyuan;Klionsky, Daniel J.
通讯作者：
Klionsky, Daniel J.