Acetylation of SCFD1 regulates SNARE complex formation and autophagosome-lysosome fusion
Acetylation of SCFD1 regulates SNARE complex formation and autophagosome-lysosome fusion
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SCFD1 乙酰化调节 SNARE 复合物形成和自噬体-溶酶体融合
DOI：
10.1080/15548627.2022.2064624
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发表时间：
2022-04-24
期刊：
影响因子：
13.3
通讯作者：
Liu, Rong
中科院分区：
文献类型：
--
作者：
Huang, Hong;Ouyang, Qinqin;Liu, Rong
摘要SCFD1(sec1家族结构域包含1)最近被证明在自噬-溶酶体融合中发挥作用，多项研究表明乙酰化(翻译后修饰)对巨型自噬/自噬的调节作用。在这里，我们证明了乙酰化和磷酸化依赖的机制控制SCFD1ʹS在自噬小体-溶酶体融合中的功能。在自噬刺激条件下检测到SCFD1乙酰化程度的降低后，我们发现KAT2B/PCAF催化SCFD1的K126和K515残基的乙酰化；我们还发现这两个残基被SIRT4去乙酰化。重要的是，我们证明了AMPK控制的SCFD1磷酸化强烈破坏了SCFD1与KAT2B相互作用的能力，从而确保SCFD1乙酰化水平保持在较低水平。最后，我们证明了SCFD1乙酰化抑制自噬通量，特别是通过阻断STX17-SNAP29-VAMP8 SNARE复合体的形成。因此，我们的研究揭示了SCFD1的磷酸化和乙酰化修饰通过介导SNAR复合体的形成来调节自噬体成熟的机制。ACLY：ATP柠檬酸裂解酶；CREB：cAMP反应元件结合蛋白；EBSS：营养剥夺介质；EP300：E1A结合蛋白p300；KAT5/Tip60：赖氨酸乙酰转移酶5；HOPS：同型融合和蛋白质分类；MS：质谱学；SCFD1：Sec1家族结构域包含1；SM：Sec1/Munc18；SNARE：可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体；UVRAG：辐射抗性相关的紫外线相关连接蛋白。
ABSTRACT SCFD1 (sec1 family domain containing 1) was recently shown to function in autophagosome-lysosome fusion, and multiple studies have demonstrated the regulatory impacts of acetylation (a post-translational modification) on macroautophagy/autophagy. Here, we demonstrate that both acetylation- and phosphorylation-dependent mechanisms control SCFD1ʹs function in autophagosome-lysosome fusion. After detecting a decrease in the extent of SCFD1 acetylation under autophagy-stimulated conditions, we found that KAT2B/PCAF catalyzes the acetylation of residues K126 and K515 of SCFD1; we also showed that these two residues are deacetylated by SIRT4. Importantly, we showed that AMPK-controlled SCFD1 phosphorylation strongly disrupts the capacity of SCFD1 to interact with KAT2B, thus ensuring that the SCFD1 acetylation level remains low. Finally, we demonstrated that SCFD1 acetylation inhibits autophagic flux, specifically by blocking STX17-SNAP29-VAMP8 SNARE complex formation. Thus, our study reveals a mechanism through which phosphorylation and acetylation modifications of SCFD1 mediate SNARE complex formation to regulate autophagosome maturation.ACLY: ATP citrate lyase; CREB: cAMP responsive element binding protein; EBSS: nutrient-deprivation medium; EP300: E1A binding protein p300; KAT5/TIP60: lysine acetyltransferase 5; HOPS: homotypic fusion and protein sorting; MS: mass spectroscopy; SCFD1: sec1 family domain containing 1; SM: Sec1/Munc18; SNARE: soluble N-ethylmaleimide-sensitive factor attachment protein receptor; UVRAG: UV radiation resistance associated