Prefused lysosomes cluster on autophagosomes regulated by VAMP8.
Prefused lysosomes cluster on autophagosomes regulated by VAMP8.
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DOI：
10.1038/s41419-021-04243-0
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发表时间：
2021-10-13
影响因子：
9
通讯作者：
Diao J
中科院分区：
文献类型：
--
作者：
Chen Q;Hao M;Wang L;Li L;Chen Y;Shao X;Tian Z;Pfuetzner RA;Zhong Q;Brunger AT;Guan JL;Diao J
溶酶体与自噬体融合是自噬体成熟的关键。虽然已经确定了几种调节这种融合过程的蛋白质，但预融合结构及其调控仍不清楚。在此，我们表明，在刺激下，多个溶酶体在单个自噬体周围形成簇，为膜融合奠定了基础。溶酶体上的可溶性n -乙基丙烯酰亚胺敏感因子附着蛋白受体（SNARE）蛋白-囊泡相关膜蛋白8 (VAMP8) -在溶酶体簇的预融合状态形成中起重要作用。为了研究磷酸化对自发融合的潜在作用，我们研究了VAMP8 c端残基磷酸化的影响。使用磷酸化模拟，我们观察到在集合脂质混合试验中融合减少，与自噬体相关的未融合溶酶体增加。这些结果表明，磷酸化不仅在正常情况下减少自发融合以使自噬通量最小化，而且还可以预先组装多个溶酶体以增加融合概率，以便在刺激时恢复自噬。因此，VAMP8磷酸化可能通过影响自噬体成熟在化疗耐药中发挥重要作用。
Lysosome–autophagosome fusion is critical to autophagosome maturation. Although several proteins that regulate this fusion process have been identified, the prefusion architecture and its regulation remain unclear. Herein, we show that upon stimulation, multiple lysosomes form clusters around individual autophagosomes, setting the stage for membrane fusion. The soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein on lysosomes—vesicle-associated membrane protein 8 (VAMP8)—plays an important role in forming this prefusion state of lysosomal clusters. To study the potential role of phosphorylation on spontaneous fusion, we investigated the effect of phosphorylation of C-terminal residues of VAMP8. Using a phosphorylation mimic, we observed a decrease of fusion in an ensemble lipid mixing assay and an increase of unfused lysosomes associated with autophagosomes. These results suggest that phosphorylation not only reduces spontaneous fusion for minimizing autophagic flux under normal conditions, but also preassembles multiple lysosomes to increase the fusion probability for resuming autophagy upon stimulation. VAMP8 phosphorylation may thus play an important role in chemotherapy drug resistance by influencing autophagosome maturation.
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影响因子：
64.8
作者：
Diao, Jiajie;Liu, Rong;Rong, Yueguang;Zhao, Minglei;Zhang, Jing;Lai, Ying;Zhou, Qiangjun;Wilz, Livia M.;Li, Jianxu;Vivona, Sandro;Pfuetzner, Richard A.;Brunger, Axel T.;Zhong, Qing
通讯作者：
Zhong, Qing
影响因子：
16.6
作者：
Hsieh, Cheng-Wei;Yang, Wei Yuan
通讯作者：
Yang, Wei Yuan
影响因子：
64.5
作者：
Green DR;Levine B
通讯作者：
Levine B
影响因子：
5
作者：
Desantis V;Saltarella I;Lamanuzzi A;Mariggiò MA;Racanelli V;Vacca A;Frassanito MA
通讯作者：
Frassanito MA
影响因子：
64.5
作者：
Delarue, M.;Brittingham, G. P.;Pfeffer, S.;Surovtsev, I. V.;Pinglay, S.;Kennedy, K. J.;Schaffer, M.;Gutierrez, J. I.;Sang, D.;Poterewicz, G.;Chung, J. K.;Plitzko, J. M.;Groves, J. T.;Jacobs-Wagner, C.;Engel, B. D.;Holt, L. J.
通讯作者：
Holt, L. J.