机构：[a]The affiliated Hospital of Guilin Medical University, Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guangxi Neurological Diseases ClinicalResearch Center, Guilin, Guangxi, China[b]Rutgers Cancer Institute of New Jersey, Rutgers University, New Brunswick, New Jersey, USA[c]Department ofPharmacy, Xijing Hospital, Fourth Military University, Xi’ an, Shanxi, China[d]Department of Radiation Oncology, The First Hospital of Jilin University,Changchun, China[e]Department of General Surgery, the Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China[f]Department ofGeneral Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China[g]Department of Gastrointestinal Surgery, ZhongshanHospital of Xiamen University, Institute of Gastrointestinal Oncology, Medical College of Xiamen University, Xiamen, Fujian, China[h]Department ofGastrointestinal Oncology, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang, China[i]Department of Clinical Medicine, West China University ofMedical Science, Chengdu, Sichuan, China[j]Department of Histology and Embryology, Xiang Ya School of Medicine, Central South University, Changsha,Hunan, China[k]Laboratory of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Guilin Medical University, Guilin, Guangxi 541001, China[l]State KeyLaboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, School of Preclinical Medicine, Xinjiang Medical University,Urumqi, Xinjiang, China[m]Department of Neurology, the affiliated hospital of Guilin Medical University, Guilin, Guangxi, China[n]Department ofGastrointestinal Surgery, Zhongshan Hospital of Xiamen University, Xiamen, Fujian, China[o]Department of Surgery, Robert-Wood-Johnson Medical SchoolUniversity Hospital, Rutgers University, The State University of New Jersey, New Brunswick, New Jersey 08901, USA
UVRAG (UV radiation resistance associated) is an important regulator of mammalian macroautophagy/autophagy by interacting with BECN1, PIK3C3, and RUBCN. Phosphorylation of UVRAG by MTORC1 negatively regulates autophagosome maturation under nutrient-enriched conditions. However, how UVRAG ubiquitination is regulated is still unknown. Here we report that UVRAG is ubiquitinated by SMURF1 at lysine residues 517 and 559, which decreases the association of UVRAG with RUBCN and promotes autophagosome maturation. However, the deubiquitinase ZRANB1 specifically cleaves SMURF1-induced K29 and K33-linked polyubiquitin chains from UVRAG, thereby increasing the binding of UVRAG to RUBCN and inhibiting autophagy flux. We also demonstrate that CSNK1A1-mediated UVRAG phosphorylation at Ser522 disrupts the binding of SMURF1 to UVRAG through PPxY motif and blocks UVRAG ubiquitination-mediated autophagosome maturation. Interestingly, ZRANB1 is phosphorylated at Thr35, and Ser209 residues by CSNK1A1, and this phosphorylation activates its deubiquitinating activity. Importantly, we provide in vitro and in vivo evidence that UVRAG ubiquitination at lysine residues 517 and 559 or prevention of Ser522 phosphorylation by D4476, a CSNK1A1 inhibitor, enhances the lysosomal degradation of EGFR, which significantly inhibits hepatocellular carcinoma (HCC) growth. Furthermore, UVRAG S522 phosphorylation levels correlate with ZRANB1 T35/S209 phosphorylation levels and poor prognosis in HCC patients. These findings identify a novel molecular mechanism by which ubiquitination and phosphorylation of UVRAG regulate its function in autophagosome maturation and HCC growth, encouraging further study of their potential therapeutic implications. Abbreviations: ATG: autophagy related; BafA1: bafilomycin A1; BECN1: beclin 1; CHX: cycloheximide; CSNK1A1/CK1α: casein kinase 1 alpha 1; CQ: chloroquine; DUB: deubiquitinase; EBSS: Earle's balanced salt solution; EGF: epidermal growth factor; GFP: green fluorescent protein; GST: glutathione S-transferase; HBSS: Hanks balanced salts solution; HCC: hepatocellular carcinoma; MAP1LC3B/LC3: microtubule associated protein 1 light chain 3 beta; MEFs: mouse embryo fibroblasts; mRFP: monomeric red fluorescent protein; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; PTMs: post-translational modifications; RUBCN: rubicon autophagy regulator; siRNA: small interfering RNA; SMURF1: SMAD specific E3 ubiquitin protein ligase 1; SQSTM1: sequestosome 1; Ub-AMC: ubiquitin-7-amido-4-methylcoumarin: a fluorogenic substrate; UVRAG: UV radiation resistance associated; ZRANB1/TRABID: zinc finger RANBP2-type containing 1.
基金：
grants from Rutgers University Pilot
Project, National Natural Science Foundation of China (81602149,
81702387, 31571241, and 31660266).
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2019]版：
大类|2 区生物学
小类|2 区细胞生物学
最新[2023]版：
大类|1 区生物学
小类|2 区细胞生物学
第一作者：
第一作者机构：[a]The affiliated Hospital of Guilin Medical University, Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guangxi Neurological Diseases ClinicalResearch Center, Guilin, Guangxi, China[b]Rutgers Cancer Institute of New Jersey, Rutgers University, New Brunswick, New Jersey, USA
共同第一作者：
通讯作者：
通讯机构：[a]The affiliated Hospital of Guilin Medical University, Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guangxi Neurological Diseases ClinicalResearch Center, Guilin, Guangxi, China[j]Department of Histology and Embryology, Xiang Ya School of Medicine, Central South University, Changsha,Hunan, China[l]State KeyLaboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, School of Preclinical Medicine, Xinjiang Medical University,Urumqi, Xinjiang, China[m]Department of Neurology, the affiliated hospital of Guilin Medical University, Guilin, Guangxi, China[n]Department ofGastrointestinal Surgery, Zhongshan Hospital of Xiamen University, Xiamen, Fujian, China[o]Department of Surgery, Robert-Wood-Johnson Medical SchoolUniversity Hospital, Rutgers University, The State University of New Jersey, New Brunswick, New Jersey 08901, USA[*a]The affiliated Hospital of GuilinMedical University, Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guangxi Neurological Diseases Clinical Research Center, Guilin, Guangxi, China[*b]Department of Gastrointestinal Surgery, Zhongshan Hospital of Xiamen University, Xiamen, Fujian, China[*c]The affiliated Hospital of Guilin Medical University, Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guangxi Neurological Diseases Clinical Research Center, Guilin, Guangxi, China[*d]Department of Histology and Embryology, Xiang Ya School of Medicine, Central South University, Changsha, Hunan, China
推荐引用方式(GB/T 7714)：
Xing Feng,Yanyan Jia,Yuyu Zhang,et al.Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth.[J].Autophagy.2019,15(7):1130-1149.doi:10.1080/15548627.2019.1570063.
APA：
Xing Feng,Yanyan Jia,Yuyu Zhang,Fei Ma,Yuekun Zhu...&Zhiyong Zhang.(2019).Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth..Autophagy,15,(7)
MLA：
Xing Feng,et al."Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth.".Autophagy 15..7(2019):1130-1149
生物学