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An 11S proteasome regulatory pathway controls anticancer sensitivity in p53-deficient tumor cells

An 11S proteasome regulatory pathway controls anticancer sen...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Xiaotao Li Shanghai Key Laboratory of Regulatory Biology Institute of Biomedical SciencesSchool of Life SciencesEast China Normal University

Ubiquitin-independent protein degradation is an emerging new field in proteasome-mediated protein *** proteasome activator,REG,is a major player mediating the ubiquitin-independent degradation of proteins including numerous tumor *** in many human cancers,REG promotes cancer progression in animal *** this REGproteasome pathway is regulated,particularly for the control of p21/CIP in p53 defective cancer cells during cell cycle or genotoxic insults,remains *** we show that NIP30,a novel REGγ regulator,inhibits REG activity,attenuates cell growth,and sensitizes p53-compromised cells to chemotherapeutic *** inhibitory role of NIP30 requires tight binding to REG via an evolutionarily conserved serine-rich domain with phosphorylation at 4 specific serine *** find the cyclindependent phosphatase,CDC25 A,regulates phosphorylation of NIP30,required for binding to REG,and therefore the REGdependent degradation of p21/CIP during the cell cycle or DNA damage *** of p21 in vivo by the CDC25 ANIP30-REG pathway was validated in p53-/-and p53/REG double-deficient *** phospho-NIP30 mimetics significantly enhance the growth inhibitory effect of anti-cancer drugs in vitro and in *** results provide insights into the regulatory signaling that controls the ubiquitin-independent REG-proteasome system in modulating *** this avenue of regulation of p53-independent cell growth may allow a promising approach in cancer therapy.

关键词： NIP An 11S proteasome regulatory pathway controls anticancer sensitivity in p53-deficient tumor cells

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How ligand-receptor binding kinetics affects downstream gene expression

How ligand-receptor binding kinetics affects downstream gene...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者：郭安南张晓玲来鲁华北京大学前沿交叉学科研究院生命科学联合中心北京大学前沿交叉学科研究院定量生物学中心

近年来研究表明,配体受体结合动力学在信号通路功能选择性方面发挥重要作用。NF-κB控制着下游与炎症、免疫应答、细胞增殖分化等生理功能相关的数百个基因的表达,其活性的错误调控将引起多种疾病。不同的TNFα-TNFR1结合动力学将会导致... 详细信息

近年来研究表明,配体受体结合动力学在信号通路功能选择性方面发挥重要作用。NF-κB控制着下游与炎症、免疫应答、细胞增殖分化等生理功能相关的数百个基因的表达,其活性的错误调控将引起多种疾病。不同的TNFα-TNFR1结合动力学将会导致NF-κB入核动力学行为发生改变,更快结合动力学情况下,NF-κB反应时间更短,半峰宽更小。利用高通量测序手段,我们发现结合动力学的不同会引起下游基因表达谱的变化,通过差异表达分析和功能注释等去研究配体受体结合动力学在功能选择性方面的作用。探索不同结合动力学的TNFα突变体来靶向调控基因表达,这在生物学研究以及医学应用领域都具有很高价值。

关键词：功能选择性结合动力学基因表达谱

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TOPK inhibits autophagy by phosphorylating ULK1 to promote chemoresistance in glioma cells

TOPK inhibits autophagy by phosphorylating ULK1 to promote c...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Hui Lu Changshu Ke Xiaofang Ni Juanjuan Xiao Ruijuan Xiu Qin Tian Huaxiong Pan Ling Zou Fei Wang Tengfei Ma Ping Yuan Lin Liu Jianmin Zhang Wei Jia Qiuhong Duan Feng Zhu Department of Biochemistry and Molecular Biology School of Basic MedicineHuazhong University of Science and Technology Department of Pathology Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology Department of Pathology Union HospitalHuazhong University of Science and Technology

TMZ(temozolomide) treatment,as first-line chemotherapy in GBM(glioblastma),faced with drug *** studies emphasize that TMZ resistance is associated with autophagy,but the mechanism remained particularly ***1,as a serine-threonine protein kinase,is the upper-most protein of ULK1 complex and is emerging as a crucial node in autophagy ***,the regulation of ULK1 is not fully ***(T-LAK cell-originated protein kinase) is a member of MAPKK protein family,which is highly expressed in many malignant tumors and can be used as a molecular marker for the diagnosis and prognosis of *** this study,we identified TOPK,an oncokinase,as a novel upstream kinase to phosphorylate ULK1,and TOPK could inhibit autophagy in glioma *** found that TOPK could directly bind with and phosphorylate ULK1 at Ser495,Ser469,and Ser533 in vitro and ex *** phosphorylation of ULK1 at Ser495,Ser469,and Ser533 by TOPK decreased the activity and stability of ***,as the decrease of ULK1 activity will reduce the occurrence of autophagy,we would like to check the induction of *** demonstrated that TOPK could inhibit the induction and progression of autophagy in glioma ***,TOPK inhibition increased the sensitivity of glioma cells to *** discovery sheds light on the problem of TMZ-resistance in GBM treatment.

关键词： TOPK ULK1 autophagy temozolomide chemoresistance

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The m6A RNA Methylation Enhances 6-phosphogluconate dehydrogenase Expression to Promote Lung Cancer Growth

The m6A RNA Methylation Enhances 6-phosphogluconate dehydrog...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Hao Sheng Yanke Guo Zhen Li Xiaoya Zhang Zhaoliang Lu Qi Feng Ronghui Liang Yanping Li Leilei Li Jing Li Dan Li Shuangping Liu Shuai Zhang Changliang Shan The First Affiliated Hospital Biomedical Translational Research InstituteGuangdong Province Key Laboratory of Molecular Immunology and Antibody EngineeringJinan University State Key Laboratory of Medicinal Chemical Biology College of Pharmacy and Tianjin Key Laboratory of Molecular Drug ResearchNankai University Division of Cellular & Molecular Research Humphrey Oei Institute of Cancer ResearchNational Cancer Centre Singapore Department of Pathology Medical SchoolDalian University Department of Medical Biochemistry and Molecular Biology School of MedicineJinan University

6-phosphogluconate dehydrogenase(6 PGD),the third enzyme in the oxidative pentose phosphate pathway(PPP) contributes to tumor *** previous study showed that lung cancer patients with higher 6 PGD levels had worse survival outcomes compared with patients with lower 6 PGD ***,how 6 PGD is upregulated in human lung cancers is *** this study,we demonstrate that 6 PGD is upregulated in lung cancer tissues and lung cancer cells relative to normal tissues and normal cells,***,we indentify methyltransferase-like 3(METTL3) as 6 PGD mRNA methyltransferase,and fat mass-and obesity-associated protein(FTO) as 6 PGD mRNA ***,we identify YTHDF2 regulates 6 PGD protein expression by facilitating its mRNA translation,which dependent on 6 PGD mRNA m6 A *** further find that YTHDF2,METTL3 and FTO would reprogram oxidative PPP as well as cancer cell proliferation and tumor growth via regulating 6 PGD *** together,our results demonstrate that m6 A modification regulates 6 PGD expression by promoting its mRNA translation and serves as a promising anticancer target.

关键词： 6-phosphogluconate dehydrogenase RNA m6A modification metabolism oxidative pentose phosphate pathway

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Novel crystal structure of dextranase from marine Arthrobacter oxydans KQ11 and improvement of its thermostability by semi-rational design B-factor

Novel crystal structure of dextranase from marine Arthrobact...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Wei Ren Le Liu Lide Gu Wanli Yan Dongxue Dong Mingsheng Lyu Shujun Wang Changhai Wang Key Laboratory of Marine Biology Nanjing Agricultural University Jiangsu Marine Resources Development Research Institute Huaihai Institute of Technology College of Marine Life and Fisheries HuaiHai Institute of Technology Co-Innovation Center of Jiangsu Marine Bio-industry Technology Huaihai Institute of Technology

Dextranases(α-1,6-D-glucan-6-glucanohydrolase;EC 3.2.1.11) hydrolyze dextran to oligosaccharides at the α-1,6 glucosidic *** structure of dextranase from marine Arthrobacter oxydans KQ11 was determined at 1.4 ? *** is the third structure of dextranase,the second structure of a family 49 glycoside hydrolase(GH49) member,and the first from marine *** crystal structure of the conserved fragment from Aal52 to Thr638 of Arthrobacter oxydans dextranase (Aodex) consists of the following two domains: the N-terminal domain A and the C-terminal domain B.N-terminal domain A is identified as a β sandwich,which is connected to a right-handed parallel β-helical at the *** results of analysis of sequence comparison,cavity region of Aodex structure surface and docking suggested that catalytic reaction mechanism of Aodex is an Asp420/Glu423 double displacement mechanism of retaining *** thermostability of S357 F was obviously improved by site-directed mutagenesis based on B-factor values,which indicates that semi-rational design B-factor method achieved superior results with a significantly reduced number of variants to be ***507 plays a role in the expulsion of cleaved dextran after the hydrolysis reaction,leading to the rapid turnover of the enzyme,and which may form aromatic-aromatic interactions contributing to the thermostability with mutant *** knowledge about the structure and the thermostability mutants of Aodex from marine bacteria provides insights into the development of strategies that will be more suitable for industrial use.

关键词： Crystal structure Dextranase from marine Arthrobacter oxydans Double displacement mechanism of retaining enzyme Thermostability Semi-rational design B-factor

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SLC27A5缺失通过激活Keap1/Nrf2通路促进肝癌增殖的机制研究

SLC27A5缺失通过激活Keap1/Nrf2通路促进肝癌增殖的机制研究

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者：高庆祝张桂冀郑亚秋杨翼黄爱龙汪凯唐霓重庆医科大学感染性疾病分子生物学教育部重点实验室

目的研究溶质载体家族27成员5(Solute Carrier Family 27 Member 5,SLC27A5)在肝癌发生发展中的作用及其分子机制。方法利用TCGA数据分析肝癌与癌旁组织中SLC27A5表达情况及其与预后的关系,并在临床肝癌标本中进行Western Blot和免疫组... 详细信息

目的研究溶质载体家族27成员5(Solute Carrier Family 27 Member 5,SLC27A5)在肝癌发生发展中的作用及其分子机制。方法利用TCGA数据分析肝癌与癌旁组织中SLC27A5表达情况及其与预后的关系,并在临床肝癌标本中进行Western Blot和免疫组化验证;利用重组腺病毒和CRISPR/Cas9系统,分别建立SLC27A5过表达和基因敲除两种细胞模型,通过MTS、EdU和克隆形成实验观察SLC27A5对肝癌细胞增殖的影响,通过裸鼠皮下成瘤和肝原位移植瘤模型观察SLC27A5对肿瘤增殖的影响;利用RNA-Seq技术分析SLC27A5调控的靶基因,q-PCR、Western Blot和IHC技术证实SLC27A5对靶基因的调控;利用抗氧化反应元件(Anti-oxidant response element, ARE)-荧光素酶活性报告系统、q-PCR和Western Blot检测Keap1/Nrf2信号通路的活化状态;利用CellROX?Orange探针,丙二醛(MDA)检测试剂盒和4-羟基壬烯酸(4-HNE)抗体分别检测细胞内活性氧(ROS)和脂质过氧化水平;利用超高效液相色谱-串联质谱/质谱法(UPLC-MS/MS)测定4-HNE对Keap1蛋白半胱氨酸残基的修饰;肝癌细胞、裸鼠皮下成瘤和肝原位移植瘤模型中,观察沉默TXNRD1联合索拉菲尼处理对肝癌细胞和肿瘤增殖和凋亡能力的影响。结果 TCGA数据分析和临床肝癌组织检测发现SLC27A5在肝癌组织中显著低表达(p<0.001),其表达与肝细胞癌患者预后呈正相关(p<0.01)。过表达SLC27A5抑制PLC/PRF/5和SK-Hep1细胞增殖;敲除SLC27A5基因促进HepG2细胞增殖,裸鼠皮下成瘤模型中过表达SLC27A5抑制肝癌增殖;肝原位移植瘤模型中敲除SLC27A5促进肝癌增殖。过表达SLC27A5下调TXNRD1表达,敲除SLC27A5上调TXNRD1的表达,临床肝癌组织标本中SLC27A5 mRNA表达与TXNRD1 mRNA表达成负相关(r=-0.35,p=0.02)。过表达SLC27A5抑制ARE荧光素酶活性、Nrf2核转位和下游靶基因表达;敲除SLC27A5增加ARE荧光素酶活性、促进Nrf2核转位和下游靶基因表达。过表达SLC27A5抑制ROS产生和脂质过氧化,敲除SLC27A5促进ROS产生和脂质过氧化。UPLC-MS/MS检测4-HNE修饰Keap1蛋白半胱氨酸残基位点是Cys513和Cys518。与单用索拉非尼相比,沉默TXNRD1联合索拉菲尼明显抑制肝癌细胞和肿瘤增殖、促进肝癌细胞和肿瘤凋亡。结论 SLC27A5异常低表达,增加ROS和脂质过氧化水平,激活Keap1/Nrf2抗氧化信号通路,上调TXNRD1表达,促进肝癌增殖。

关键词： SLC27A5 肝细胞癌 ROS Keap1/Nrf2信号通路 TXNRD1 索拉非尼

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Linking sno-lncRNA and SPA lncRNAs to Prader-Willi syndrome

Linking sno-lncRNA and SPA lncRNAs to Prader-Willi syndrome

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Huang Wu Qing-Fei Yin Zheng Luo Li Yang Ling-Ling Chen State Key Laboratory of Molecular Biology CAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of Sciences Key Laboratory of Computational Biology Shanghai CAS-MPG Partner Institute for Computational BiologyCAS Center for Excellence in Brain Science and Intelligence TechnologyChinese Academy of Sciences School of Life Science ShanghaiTech University

Prader-Willi syndrome(PWS) is a neuro-developmental genetic disorder characterized with obesity and *** cases of PWS can be attributed to a large(5-6 Mb) paternal deletion of chr15 q11-q13,but the minimal deletion is mapped to the 3'-untranslated region(UTR) of SNURF-SNRPN that expresses dozens of snoRNAs and several lncRNAs including sno-lncRNAs and *** PWS region sno-lncRNAs and SPAs are processed by distinct ***-lncRNAs are derived from introns and processed on both ends by snoRNP machinery,leading to the accumulation of lncRNAs flanked by snoRNA sequences lacking 5'-m7 G caps and 3'-poly(A) *** processing is associated with nascent mRNA 3' processing and kinetic competition between XRN2 trimming and Pol II *** cleavage/polyadenylation of its upstream gene,the downstream uncapped pre-SPA is trimmed by XRN2 until this exonuclease reaches the co-transcriptionally assembled *** prevents further degradation,and allows continuous Pol II *** lncRNAs form a nuclear accumulation that is enriched in RBPs including TDP43,RBFOX2,and *** of a human PWS embryonic stem cells cellular model by depleting these lncRNAs resulted in altered patterns of RBPs binding and alternative *** of normal and PWS-hESCs to hypothalamic-like neurons showed remarkable gene expression changes,including those have reported functions in controlling of appetites and *** findings suggest that the absence of these sno-processed lncRNAs in PWS patients is linked to PWS pathogenesis.

关键词： PWS sno-lncRNA SPA Alternative splicing Hypothalamic-like neurons

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RhTyrRS(Y341A), a Novel Human Tyrosyl-tRNA Synthetase Mutation, Stimulate Thrombopoietic Through Activation of VEGF-R Ⅱ//NF-κB Pathway

RhTyrRS(Y341A), a Novel Human Tyrosyl-tRNA Synthetase Mutati...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Shi yun Zhang Yanling Wangxu Zhan Ling Yu Min Mo Wei The Key Laboratory of Metabolism and Molecular Medicine Ministry of EducationFudan University The Department of Biochemistry and Molecular Biology School of Basic Medical Sciences and Institutes of Biomedical SciencesFudan University

Tumor chemo-and radiotherapy induces hematopoietic organ damage and reduces thrombocyte *** platelet transfusion strategies or drug therapies are used to treat ***,there may cause a series of side effects,such as infectious diseases and TPO ***,a more secure strategy should be explored to overcome and compensate for the shortcomings of conventional *** studies have shown that tRNA synthetase plays not only catalytic tRNA aminoacylation roles,but also functions similarly to *** previous work show that recombinant human tyrosyl-tRNA synthetase with a mutated Y341 A(tyrosine mutated to alanine)(rhTyrRS(Y341 A)) promotes megakaryocyte migrate from bone marrow to peripheral *** we indicated that rhTyrRS(Y341 A) would promote megakaryocytes in the lungs adhering to pulmonary vascular endothelial cells(PVECs) and resulting in the platelet ***,the possible mechanisms of thrombopoietic activity remain ***,we found that both of the expression of vascular cell adhesion molecule(VCAM-1) and the phosphorylation level of vascular endothelial growth factor receptor 2(VEGF-R Ⅱ) in the PVECs of rats treated with rhTyrRS(Y341 A) were significantly increased,meanwhile,the transcription factor NF-κB which play a significant role in the upregulation of the VCAM-1 gene were activated in *** addition,we also found the same results in human umbilical vein endothelial cells(HUVECs) treated with rhTyrRS(Y341 A).Double luciferase report experiment excluded the possible direct interaction between NF-κB and ***,the results provided evidence for that rhTyrRS(Y341 A) activate NF-κB to upregulate VCAM-1 in a VEGF-R Ⅱ/NF-κB pathway-dependent,resulting in megakaryocyte migrate from bone marrow to PVECs to platelet production.

关键词： VCAM-1 rhTyrRS(Y341A) NF-κB VEGF-R Ⅱ HUVECs

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Overexpression of Sirt7 Promotes Human Colorectal Cancer Cell Colony Formation Through Inhibiting SESN2-mediated Autophagy

Overexpression of Sirt7 Promotes Human Colorectal Cancer Cel...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Yang Li Shun Zhang Dan-dan Huang Jing Lin Ting Cai Ningbo No.2 Hospital

Background: Sirt7,a member of the sirtuin family,deacetylates Histone H3 K18 ac thus maintaining oncogenic *** less is known about the relationship between colorectal cancer development and *** we explore the role of Sirt7 in colorectal cancer cell autophagy and colony ***: Expression of Sirt7 protein and mRNA in human colorectal cancer specimens and cell lines were assessed using immunohistochemistry and *** effects of Sirt7 expression on colony formation and autophagy was evaluated after shRNA *** levels of autophagy related proteins(Beclin1,ATG5,ATG7,SESN2 and p62) were also *** upstream regulators of SESN2 were determined using ChIP assay and its effect on autophagy and colony formation was further verified after shRNA knockdown in colorectal cancer ***: Sirt7 was markedly and reproducibly upregulated in human colorectal cancers and their cell-line *** of Sirt7 in human colorectal cancer cell lines HCT116 and SW620 dramatically reduced their colony *** analyses showed that Sirt7 overexpression inhibited SESN2-mediated-autophagy and further promoted human colorectal cancer cell colony ***,Sirt7 overexpression decreased Sp1 protein expression and further reduced Sp1 interaction with SESN2 promoter,leading to the decrease of SESN2 transcription ***: Our results provide the mechanistic link between autophagy and human colorectal cancer cell colony *** signaling axis along Sirt7-Sp1-SESN2-autophagy that is highly operative in colorectal cancer cells might be explored further for developing novel diagnostic and therapeutic approaches to manage advanced colorectal cancers.

关键词： Colorectal Cancer Sirt7 SESN2 Sp1 Autophagy

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An engineered PD-1-based and MMP-2/9-oriented fusion protein exerts potent antitumor efficacy against melanoma

An engineered PD-1-based and MMP-2/9-oriented fusion protein...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Mulan Wei Xujie Liu Chunyu Cao Jianlin Yang Yafeng Lv Jiaojiao Huang Yanlin Wang Ye Qin Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy Three Gorges University Medical College Department of Pathology Yiling Hospital Yichang Institute of Medicinal Biotechnology Chinese Academy of Medical Sciences and Peking Union Medical College

Recent studies have shown that PD-1/PD-L1 checkpoint blockade is a dramatic therapy for melanoma through enhancing antitumor immune ***,major strategies for PD-1/PD-L1 blockade have mainly focused on the use of antibodies and *** seeking an alternative approach,it is of interest to employ endogenous proteins as blocking *** the facts that extracellular domain of PD-1(ePD1) includes the specific binding site with PD-L1,we constructed a PD-1-based recombinantly tailored fusion protein(dFv-ePD1) which consists of bivalent variable fragment(dFv) of MMP-2/9-targeted antibody and ***,the melanoma-binding intensity and antitumor activity were *** shown,intense and selective binding capability of the protein dFv-ePD1 to human melanoma specimens was confirmed by tissue *** addition,dFv-ePD1 suppressed significantly the migration and invasion of mouse melanoma cells B16-F1 and displayed the cytotoxicity to cancer cells in ***,dFv-ePD1 significantly inhibited the growth of mouse melanoma B16-F1 tumor in mice and in vivo fluorescence imaging showed that dFv-ePD was gradually accumulated in B16-F1 tumor,whose fluorescence intensity at tumor site was stronger than that of *** study indicates that the recombinant protein dFv-ePD1 have an intensive melanoma-binding capability and exert potent therapeutic efficacy against *** novel format of PD-L1-blocked agent may play an active role in antitumor immunotherapy.

关键词： PD-1/PD-L1 MMP-2/9 melanoma targeting protein antitumor efficacy

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