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检索条件"任意字段=中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议"

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RNF168-mediated H2A neddylation antagonizes its ubiquitinationand regulates DNA damage repair

RNF168-mediated H2A neddylation antagonizes its ubiquitinati...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Junhong Guan Tingting Li Xiaofeng Zheng Peking University

NEDD8(neural precursor cell expressed, developmentally downregulated 8), an ubiquitinlike protein, plays important roles in many biological processes including DNA damage responsepathway. However, the substrates of neddylation in this pathway are remainedlargely unknown and the relationship between ubiquitin and NEDD8 is also a very interestingquestion needed to be answered. Here, we showed that H2 A is a substrate of NEDD8,and H2 A neddylation antagonizes its ubiquitination. Up-regulation of the free NEDD8 suppressesH2 A ubiquitination and decrease of the free NEDD8 level promotes H2 A *** further found that an E3 ligase RNF168 promotes both H2 A ubiquitination andneddylation. Interestingly, NEDD8 also targets RNF168 and regulates its E3 ubiquitin *** of RNF168 neddylation impairs the interaction between RNF168 and its E2Ubc13. Furthermore, in response to DNA damage, H2 A neddylation decreases with an increaseof H2 A ubiquitination which facilitates DNA damage repair. And at the late stage ofdamage repair, H2 A neddylation increases gradually while ubiquitination decreases to thebasal levels. Mechanistically, NEDD8 negatively regulates DNA damage repair process bysuppressing the ubiquitination of H2 A and γH2AX, which further blocks the recruitment ofdamage-response protein BRCA1. Our findings elucidate the relation of H2 A ubiquitinationand neddylation, and suggest a novel modulate approach to DNA damage repair throughthe neddylation pathway.

关键词： Neddylation DNA damage response H2A

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MiR-143 inhibits cell proliferation by targeting ATG2B in non-smallcell lung cancer H1299 cells

MiR-143 inhibits cell proliferation by targeting ATG2B in no...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： jiali wei zhongliang ma yanli li botao zhao detao wang yan jin youxin jin Schoolof life sciences Shanghai University School of Life Sciences Shanghai University Shenzhen Institutes of advanced Technology Chinese Academy of Sciences State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Institutes for Biological SciencesChinese Academy of Sciences

miRNAs are small non-coding RNAs involving in multiple biological pathways by posttranscriptionalregulation of gene expression. Recently, autophagy was reported to suppressprogression of non-small cell lung cancer(NSCLC). However,little is known abouthow miRNAs regulate autophagy in NSCLC. Here, our results identified the autophagygene Autophagyrelated-2b(ATG2B) as a novel target of miR-143. The overexpression ofmiR-143 could down-regulate the expression of atg2 b at both the transcriptional andtranslational levels by direct binding to its 3' untranslated region. Cell proliferation was significantlyinhibited by the ectopic expression of miR-143 in H1299 cells. Knockdown ofATG2 B resulted in similar phenotype with the miR-143 overexpression in NSCLC cells. Furthermore,knockdown of ATG2 B and hexokinase 2(HK2), a key enzyme of glycolysis andanother target of miR-143, coordinated to inhibit the proliferation of H1299 cells. Our resultsdemonstrated that miR-143 was a novel and important regulator of autophagy by targeting ATG2 B and repression of the gene expression in autophagy and high glycolysis hada coordinate effect in H1299 cells. Our study suggested that the ATG2 B might be a newpotential the rapeutic target for NSCLC. Further, it was implied that interrupting both autophagy and glycolsis would improve NSCLC therapy.

关键词： MiR-143 ATG2B miRNA NSCLC autophagy glycolysis

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A novel genome defense mechanism of non-coding DNAs againstexogenous DNAs/RNAs as inspired by social insect colony

A novel genome defense mechanism of non-coding DNAs againste...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Guo-Hua QIU Xiaojin Xie Yuru Deng Linhong Deng Changzhou University National University of Singapore

Genome sequence comparisons have revealed that in multicellular organisms the numberof protein-coding sequences is relatively constant across species, but the number of noncodingDNAs increases considerably as increasing complexity of the organism. Althoughthe function of individual non-coding DNA has been noticed recently, the reasons thatgenome needs to carry a great number of non-coding DNAs remain a mystery. Consideringthat genome must require a defense mechanism against external perturbations tomaintain its integrity and that there is a need for a more suitable metaphor to representhuman genome, we hypothesize that the abundant non-coding DNAs function collectivelyto defend genome against exogenous DNAs/RNAs via both chemical and physicalstrategies and introduce the "social insect colony" metaphor for human genome. Thishypothesis is inspired by and based on the resemblance of the structure and dynamics ofgenome to that of social insect colony. Typical social insect colony contains one or a fewqueens being protected by plentiful workers. Likewise, genome(equivalent to social insectcolony) and the small number of protein-coding sequences(queens) are protected byplentiful non-coding DNAs(workers) at cytosolic and nuclear levels. In cytosol, non-codingDNA-generated miRNA, siRNA and/or piRNA degrade exogenous DNAs/RNAs. In nucleus,non-coding DNAs together with others that locate in spatially peripheral chromatins physicallyprotect the gene-rich or transcriptionally active regions located in the central chromatinsfrom being damaged by exogenous DNAs. In addition, mobile transposable elementsof non-coding DNAs are proposed to remove the integrated exogenous DNAs. This new hypothesis appears to be supported by the experimental data that can be found in the literature.

关键词： non-coding sequences transposable element antiviral small RNA genomep rotection

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Prognostic significance of p21-activated kinase 6 expression inpatients with clear-cell renal cell carcinoma

Prognostic significance of p21-activated kinase 6 expression...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：刘维斯潘登仵倩杨柳徐洁杰顾建新复旦大学

Purpose:The object of this study was to investigate prognostic values of intratumoral p21-activated kinase 6(Pak6)expression in patients with clear-cell renal cell carcinoma(ccRCC).Methods:The immunohistochemistry in tissue microarray was used to evaluate the expressionof Pak6 in 104 patients had undergone nephrectomy at Zhongshan Hospital of *** value and clinical outcomes were ***:Intratumoral Pak6 expression was significantly lower than non-tumoral tissues(P<0.001).Moreover,Pak6 low-expression was associated with unfavorable overall survival(OS)(P=0.001)and recurrence-free survival(RFS)(P=0.001).Meanwhile,in the subgroupof TNM stage I+II,the patients with Pak6 low-expression were prone to have poor OS(P=0.014)and RFS(P=0.037).Furthermore,intratumoral Pak6 low-expression,which was correlatedwith tumor size(P=0.001)and TNM stage(P=0.006),was identified as an independentprognostic factor for OS(HR 4.109,P=0.002)and RFS(HR 3.175,P=0.002).ExtendedTNM stage system with intratumoral Pak6 expression showed a better prognosticvalue for OS(AUC0.790,P=0.022)and RFS(AUC 0.769,P=0.040).And the c-index of aCox-based model combined with ECOG-PS,TNM stage,Fuhrman grade and Pak6 is 0.83 inOS and 0.80 in ***:Intratumoral Pak6 could be a potential prognosticator for overall and recurrence-free survival in ccRCC patients after *** external validation andfunctional analysis should be pursued to assess its potential prognostic and therapeuticvalues for ccRCC patients.

关键词： ccRCC Pak6 overall survival recurrence-free survival prognosis

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Preparation of Sequence-controlled Triblock-copolymer GraftedSilica Microparticles by Sequential-ATRP for Highly EfficientGlycopeptides Enrichment and Identification by Mass Spectrometry

Preparation of Sequence-controlled Triblock-copolymer Grafte...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Weijie Qin Xiaohong Qian Beijing Proteome Research Center

As one of the most important sub-proteomes in eukaryote cells, N-glycoproteins play crucialroles in various of biological processes and have long been considered closely correlatedwith the occurrence, progression and metastasis of cancer. Comprehensive characterizationof protein N-glycosylation and association of their aberrant patterns to the correspondingcancer stage may provide a unique way to discover new diagnostic biomarkersand therapeutic drug targets. However, the extremely complex nature of biologicalsamples and relatively low abundance of N-glycosylated proteins makes the enrichment ofglycoprotein/glycopeptide a prerequisite for large scale N-glycosylation identification. Inthis work, we prepared sequence controlled triblock-copolymer grafted silicamicroparticles(TGP-SMs) by sequential atom transfer radical polymerization(sequential-ATRP) of monosaccharides and zwitterionic-ion monomers for highly efficient and selectiveglycopeptides enrichment. The triblock-copolymer is composed of sequence definedpoly-zwitterionic-ion, poly-N-acetylglucosamine and poly-mannose blocks. The glycopolymerblocks carrying densely packed pending sugars are excellent mimics of the naturalcarbohydrate clusters and may induce multivalent carbohydrate-carbohydrate interaction(CCI) with the target glycopeptides. Therefore, increased retention of glycopeptides can beexpected by the combination of CCI and zwitterionic-HILIC interaction. As a result, 1244 glycopeptides were identified after TGP-SMs enrichment from mouse liver, which are 65-120% higher than that obtained by homo-glycopolymer or random-copolymer graftedsilica microparticles prepared using the conventional free radical polymerization. Theseresults demonstrate the critical role of sequence-defined block-copolymer of TGP-SMs forobtaining enhanced affinity towards glycopeptides and the potential of this sequentialATRP strategy to integrate different affinity moieties into one enrichment material toachieve deeper coverage in

关键词： Protein-PTM N-glycosylation Enrichment Identification Mass-spectrometry

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Mammalian sterile 20-like kinase 1/2 inhibit Wnt/γ-cateninsignaling pathway by directly binding Casein kinase1 epsilon

Mammalian sterile 20-like kinase 1/2 inhibit Wnt/γ-catenins...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：许飞复旦大学上海基础医学院

The Hippo signalling pathway can suppress the Wnt/β-catenin signalling pathway through the last downstream effectors YAP( Yes- associated pro-tein)/TAZ(tafazzin).MST(mammalian sterile 20-like kinase)1 functions as the upstreamkinase of the Hippo pathway, and CK1ε(casein kinase 1ε) plays roles in the up-stream signaltransduction of the Wnt/β-catenin pathway. In the present study, using tandem afnitypurication and MS analysis, CK1ε was identied as a novel partner ofMST1. Further analysisshowed that the interaction between MST1 and CK1ε was mediated by their kinase domainsand enhanced by the activation of *** exclude the interference of the phosphorylatedYAP/TAZ,the transduction from MST1 to YAP/TAZ was blocked using anti-WW45 *** the sh-WW45 cells,MST1 still inhibited the Wnt3A-induced phosphorylationof DVL2(dishevelled2) and Wnt/β-catenin signalling by disturbing the interaction ofDVL2 and CK1ε. The growth-suppressive effect of MST1 in the presence of Wnt3 A was effectivelyrelieved by the downstream activation of the Wnt/β-catenin pathway. Moreover,MST2, the close homologue of MST1, also displayed the similar function in suppressing theWnt/β-catenin pathway. Therefore the results of the present study revealed that, inadditionto the phosphorylated YAP/TAZ, the Hippo pathway can suppress the Wnt/β-cateninpathway directly through MST1/2.

关键词： Hippo signaling pathway Wnt/β-catenin signaling pathway MST1/2 CK1ε proteini nteraction negative regulation

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Effect of IQGAP1 Knockdown on Homotypic Cell-Cell Adhesion ofEsophageal Carcinoma Cells

Effect of IQGAP1 Knockdown on Homotypic Cell-Cell Adhesion o...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Kang WANG Chun HE Xiao-Xia WANG Department of Biochemistry and Molecular Biology Shanxi Medical University

Objective: To investigate the effect of IQGAP1 knockdown by RNA interference(RNAi) onhomotypic cell-cell adhesion ofesophageal carcinoma(EC) ***: Western blot analysis was adopted to detect the protein expression of IQGAP1 inKYSE150 and EC9706 cells. Slow aggregation assay and cell dissociation assay were performedto detect the homotypic cell-cell adhesion properties of tumor cells. Further,KYSE150 and EC9706 cells were transfected with RNAi plasmid for targeting IQGAP1 geneor negative control, and then the cell lines of IQGAP1 knockdown were screened withG418. Levels of IQGAP1 protein in interference group and control group were measured byWestern blot, and the changes of homotypic cell-cell adhesion were observed by slow aggregationassay and cell dissociation ***: Compared with EC9706 cells, level of IQGAP1 protein in KYSE150 cells was lower,while the homotypic cell-cell adhesion in KYSE150 cells was stronger. Further, Westernblotting analysis showed that IQGAP1 protein expression was efficiently downregulated bystable transfection with IQGAP1 siRNA expression vector. The homotypic cell-cell adhesion was obviously enhanced by IQGAP1 ***: These data suggest that downregulation of IQGAP1 expression could significantly enhance homotypic cell-cell adhesion and decrease malignant phenotype of EC cells.

关键词： IQGAP1 homotypic cell-cell adhesion RNAi esophageal carcinoma

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Human Gdnf cDNA Knock-in Beta-casein Locus in Bovine FetalFibroblast Cells

Human Gdnf cDNA Knock-in Beta-casein Locus in Bovine FetalFi...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Fei Yu Xueming Zhang Baotou Medical College Inner Mongolia University of Science and Technology

The highlight has been given to the recombinant human proteins produced by the mammaryglands of genetically modified transgenic livestock mammals in the top field of biologicaltechnology. However, for random integration of mammary gland bioreactor, insertedgene expression level varied and stayed at a low level because of affecting by flankingDNA sequences. In contrast with random integration, site-specific integration of mammarygland bioreactor created by gene targeting at high expressing locus can remarkably improvegene expression. Glial cell line-derived neurotrophic factor(GDNF) is a novel type ofneurotrophic factor cloned in 1993. Researches revealed that it may have potential applicationin the treatment of Parkinson's disease and other diseases of the central nervoussystem as a survival factor for central dopaminergic neurons. In the presentstudy, we constructeda gene targeting vector for the human gdnf gene knock-in at the bovine betacaseingene locus so that human GDNF protein can be produced at high level in the mammarygland of the gene-targeted bovine. In order to analyze the bioactivity of the vector,the plasmid DNA was transfected into human mammary tumor cell line Bcap-37 by *** transcription polymerase chain reaction and Western-blotting analysisshowed that the transfected cells produced human GDNF mRNA and protein. The resultsshow that the constructed targeting vector pNRTCNbG has bioactivity to efficiently expressGDNF in mammary gland cells. Then,3.2×107 bovine fetal fibroblast cells were transfectedwith linearized pNRTCNbG by electroporation. After selection with G418 for 8-10 days,total 3057 cell clones were obtained. After eliminating the cell clones expressed red fluorescentprotein under a fluorescent microscope, 773 cell clones were harvested, in which 5cell clones involved in gene targeting events were conformed by PCR analysis and PCRproduct sequencing..Using the gene-targeted cells as nuclear donor, a total of 134 reconstructe

关键词： gene targeting somatic cell nuclear transfer(SCNT) GDNF beta-casein locus

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PRMT5 enhances the generation of induced pluripotent stemcells(iPSCs) from dairy goat embryonic fibroblasts(GEF) through down regulation of P53

PRMT5 enhances the generation of induced pluripotent stemcel...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Zhili Chu Bowen Niu Haijing Zhu Xin He Chunling Bai Guangpeng Li JinlianHua College of Veterinary Medicine Shaanxi Centre of Stem Cells Engineering &Technology Northwest A&F University Key Laboratory for Mammalian Reproductive Biology and Biotechnology Ministry of Education Inner Mongolia University

PRMT5, protein arginine methyltransferase 5 is thought to play a role in epigenetic reprogrammingin mouse germ cells. However, there were little information on the expressionprofile of PRMT5 and their effects on the generation of livestock induced pluripotent stemcells(iPSCs) from somatic cells. Here, we explored the PRMT5 expression profiles in dairygoat and its effects on the derivation of iPSCs from dairy goat embryonic fibroblasts(GEF).The results showed that PRMT5 is a conservative gene and were expressed widely in varioustissues and different ages' testes. PRMT5 overexpression in combination withOCT3/4, SOX2, KLF4 and cMYC(POSKM) could significantly increased the number of APpositive iPSlike colonies derived-GEF compared with OSKM alone in dairy goat. Moreover,our results demonstrated that PRMT5 overexpression stimulated GEF proliferation, anddown-regulated P53, P21(a target genes of p53) and apoptotic markerCapase3 to enhancesthe somatic cell reprogramming. This study provides an efficient model to study the mechanisms on goat somatic cell reprogramming and differentiation in the future.

关键词： embryonic stem cells(ESCs) Induced pluripotent stem cells(iPSCs) Proteina rginine methyl transferase 5(PRMT5) Pluripotency dairy goat

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Epigenome-Wide Scan Identifies Differential DNA Methylation in Schizophrenia

Epigenome-Wide Scan Identifies Differential DNA Methylation ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Rongrong Han Tingfu Du Qi Xu Institute of Basic Medical Sciences Chinese Academy of Medical Sciences

Schizophrenia(SCZ) is a severe psychiatric disease with a lifetime morbidity rate of 0.5-1.0%, which affects about 1% of the world's population. Increased understanding about thefunctional complexity of the genome has led to growing recognition about the role of epigeneticvariation in the etiology of schizophrenia.A total of 40 healthy controls 40 drug-free patients who were diagnosed with first-episodeschizophrenia, fulfilled 1-yeartreatment with risperidone and achieved remission were enrolledin our study. We profiled the DNA methylation level of whole blood DNA both beforeand after the risperidone ***-wide methylation analysis was carriedout using the Illumina Infinium HumanMethylation450 BeadChip platform, an oligonucleotidearray that interrogates > 485,000 CpG dinucleotides per sample at singlenucleotideresolution. Following bisulfite treatment of DNA using the Zymo EZ DNAmethylation kit, subsequent steps for the HumanMethylation450 BeadChip analysis werecarried out according to the manufacturer's instructions. The raw intensity of BeadChipswas scanned and extracted using BeadScan in GenomeStudio *** found a total of 938 differentially methylated CpG sites( delta β>10%;p<0.001)between healthy controls and schizophrenia patients before risperidone treatments. Therewere 24 differentially methylated CpG sites( delta β>10%;p<0.001) between schizophreniapatients before and after risperidone treatments. 20 CpG sites covered 10 genes werecommon between these two cohorts were identied, among which SNCAIP had the mostsignificant *** DNA methylation in SCZ was identied at numerous loci across the whole *** ndings support the notion that altered DNA methylation could be involved in thepathophysiology of SCZ. SNCAIP gene encodes a protein containing several protein-protein interaction domains. The encoded protein interacts with alpha-synuclein inneuronal tissue and may play a role in the formation of cytoplasmic inclusions and n

关键词： DNA Methylation Schizophrenia

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