STYK1 promotes autophagy through enhancing the assembly of autophagy-specific class III phosphatidylinositol 3-kinase complex I
STYK1 promotes autophagy through enhancing the assembly of autophagy-specific class III phosphatidylinositol 3-kinase complex I
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STYK1 通过增强自噬特异性 III 类磷脂酰肌醇 3-激酶复合物 I 的组装来促进自噬
DOI：
10.1080/15548627.2019.1687212
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发表时间：
2020
期刊：
影响因子：
13.3
通讯作者：
Tang Jingfeng
中科院分区：
文献类型：
--
作者：
Zhou Cefan;Qian Xuehong;Hu Miao;Zhang Rui;Liu Nanxi;Huang Yuan;Yang Jing;Zhang Juan;Bai Hua;Yang Yuyan;Wang Yefu;Ali Declan;Michalak Marek;Chen Xing Zhen;Tang Jingfeng
摘要巨自噬/自噬在发育、肿瘤发生以及心血管和代谢性疾病中起着关键作用。自噬特异的III类磷脂酰肌醇3-激酶复合体I(PtdIns3K-C1)对自噬小体的形成是必不可少的。然而，这种复杂结构的形成规律还需要进一步的研究。在这里，我们发现STYK1(丝氨酸/苏氨酸/酪氨酸激酶1)是受体酪氨酸激酶(RTKs)家族的成员，是一种新的自噬上游调节因子。我们发现STYK1促进了人类细胞和斑马鱼的自噬小体的形成，其特征是通过几个标记蛋白，如ATG14，WIPI1和ZFYVE1，升高了Lc3-II，降低了SQSTM1/p62的水平，增加了点状结构的形成。此外，我们观察到STYK1以同源二聚体的形式直接与PtdIns3K-C1复合体结合。酪氨酸191的磷酸化促进了与该复合体的结合，从而提高了STYK1的激酶活性。我们还证明了STYK1促进了BECN1的丝氨酸磷酸化，从而减少了BECN1和BCL2之间的相互作用。此外，我们发现STYK1优先促进了PtdIns3K-C1复合体的组装，并且是PtdIns3K-C1复合体活性所必需的。综上所述，我们的发现为自噬诱导提供了新的见解，并揭示了RTK信号转导和自噬之间存在新的串扰的证据。缩写：AICAR：5-氨基咪唑-4-甲酰胺核糖核苷酸；AMPK：腺苷5‘-单磷酸(AMP)激活的蛋白激酶；ATG：自噬相关；ATP：三磷酸腺苷；bcl2：BCL2细胞凋亡调节因子；BECN1：BECLIN 1；BRE A：灯盏花素A；Co-IP：免疫共沉淀；CRISPR：成簇的规则间隔的短回文重复序列；DAPI：4’，6-二氨基-2-苯基吲哚；EBSS：厄尔平衡溶液；GAPDH：甘油醛-3-磷酸脱氢酶；GFP：绿色荧光蛋白；GSEA：基因集分析；MAP1LC3/LC3，微管相关蛋白1轻链；3MAPK8/JNK1：丝裂原激活蛋白激酶8；MRFP：单体红色荧光蛋白；MTOR：雷帕霉素激酶的机制靶点；四甲基偶氮唑蓝：3-(4，5-二甲基噻唑-2-基)-2，5-二苯基四氮唑溴化铵；PIK3C3：磷脂酰肌醇3-激酶催化亚基类型3；PIK3R4：磷脂酰肌醇-3-激酶调节亚基4；qRT-PCR：定量逆转录聚合酶链式反应；RACK1：活化C激酶1受体；RUBCN：Rubice自噬调节因子；siRNA：小干扰；SQSTM1：隔离1；YSTK1/NOK：丝氨酸/苏氨酸/酪氨酸激酶1；TCGA：癌症基因组图谱；Ub：生物素；ULK1：UNC-51样自噬激活蛋白1；UVRAG：紫外线辐射抗性相关；WIPI1：WD重复结构域，磷酸肌醇相互作用1；ZFYVE1：锌指FYVE-type包含1
ABSTRACT Macroautophagy/autophagy plays key roles in development, oncogenesis, and cardiovascular and metabolic diseases. Autophagy-specific class III phosphatidylinositol 3-kinase complex I (PtdIns3K-C1) is essential for autophagosome formation. However, the regulation of this complex formation requires further investigation. Here, we discovered that STYK1 (serine/threonine/tyrosine kinase 1), a member of the receptor tyrosine kinases (RTKs) family, is a new upstream regulator of autophagy. We discovered that STYK1 facilitated autophagosome formation in human cells and zebrafish, which was characterized by elevated LC3-II and lowered SQSTM1/p62 levels and increased puncta formation by several marker proteins, such as ATG14, WIPI1, and ZFYVE1. Moreover, we observed that STYK1 directly binds to the PtdIns3K-C1 complex as a homodimer. The binding with this complex was promoted by Tyr191 phosphorylation, by means of which the kinase activity of STYK1 was elevated. We also demonstrated that STYK1 elevated the serine phosphorylation of BECN1, thereby decreasing the interaction between BECN1 and BCL2. Furthermore, we found that STYK1 preferentially facilitated the assembly of the PtdIns3K-C1 complex and was required for PtdIns3K-C1 complex kinase activity. Taken together, our findings provide new insights into autophagy induction and reveal evidence of novel crosstalk between the components of RTK signaling and autophagy. Abbreviations: AICAR: 5-aminoimidazole-4-carboxamide ribonucleotide; AMPK: adenosine 5‘-monophosphate (AMP)-activated protein kinase; ATG: autophagy related; ATP: adenosine triphosphate; BCL2: BCL2 apoptosis regulator; BECN1: beclin 1; Bre A: brefeldin A; Co-IP: co-immunoprecipitation; CRISPR: clustered regularly interspaced short palindromic repeats; DAPI: 4′,6-diamidino-2-phenylindole; EBSS: Earle’s balanced salt solution; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; GSEA: gene set enrichment analysis; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; MAPK8/JNK1: mitogen-activated protein kinase 8; mRFP: monomeric red fluorescent protein; MTOR: mechanistic target of rapamycin kinase; MTT: 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PIK3R4: phosphoinositide-3-kinase regulatory subunit 4; qRT-PCR: quantitative reverse transcription PCR; RACK1: receptor for activated C kinase 1; RUBCN: rubicon autophagy regulator; siRNA: small interfering RNA; SQSTM1: sequestosome 1; STYK1/NOK: serine/threonine/tyrosine kinase 1; TCGA: The Cancer Genome Atlas; Ub: ubiquitin; ULK1: unc-51 like autophagy activating kinase 1; UVRAG: UV radiation resistance associated; WIPI1: WD repeat domain, phosphoinositide interacting 1; ZFYVE1: zinc finger FYVE-type containing 1