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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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Vertebrate pore-forming toxin in innate immunity

Vertebrate pore-forming toxin in innate immunity

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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： Yun Zhang Yang Xiang Chao Yan Kunming Institute of Zoology Chinese Academy of Sciences

Vertebrate pore-forming toxin in innate immunityYang Xiang,Chao Yan,Xiao-Long Guo,Kai-Feng Zhou,Sheng-An Li,Qian Gao,Xuan Wang,Feng Zhao,Jie Liu,Wen-Hui Lee,and Yun Zhang*Key Laboratory of Animal Models and Human Disease Mechanisms of The ChineseAcademy of Sciences&Yunnan Province,Kunming Institute of Zoology,the ChineseAcademy of Sciences,Kunming,Yunnan 650223,China,*email:zhangy@*** Pore-forming toxins are a common type of bacterial toxins and are important bacterial *** is produced by Aeromonas *** is interesting that aerolysin-like proteins are also found in ***,the physiological roles of theseproteins are still *** frog species Bombina maxima lives in very harsh environments,such as pools containing microorganism-rich mud,and its skin is very"toxic."A72-kDa complex of bacterial pore-forming toxin(PFT)aerolysin-like protein(alphasubunit)and trefoil factor(TFF)(beta-subunit)with a non-covalently linked form of alphabeta2was recently identified in the skin secretions of this *** protein complexwas hence named betagamma-CAT to reflect its domain *** the present study,betagamma-CAT was found to significantly accelerate bacterial clearance,thus reducingthe mortality rate in *** frog and mouse peritonitis *** rapid maturationand release of IL-1beta triggered by betagamma-CAT were detected both in vivo and invitro and may have resulted from the oligomerization of and pore formation by the proteinwithin cellular *** protein complex should thus play an important role in initiatingand regulating immune responses to eliminate microbial *** findings couldserve as clues in investigating the possible occurrence of similar antimicrobial strategies,the effective molecules and the action mechanisms involved in *** studieswill eventually help to illustrate human pathophysiological mechanisms and develop noveltherapeutics for related diseases.

关键词： innate immunity pore-forming toxin

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Fun30, a chromatin remodeler, interacting with Wor1, functions inwhite-opaque switch of Candida albicans

Fun30, a chromatin remodeler, interacting with Wor1, functio...

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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：高宁王芳中科院生化细胞所

Candida albicans is an opportunistic pathogen, often causing both mucosal and systemicinfections in immune-compromised patients. The C. albicans can switch heritably and athigh frequency between several phenotypes, including white-opaque switching. Wor1 is amaster regulator of white-opaque switching in C. albicans and is required for opaque cellformation. In order to study the regulatory network of Wor1 in white-opaque switch, weapplied yeast two hybrid system to screen the *** genomic library for Wor1 interactingproteins. Using LexA-Wor1 as bait, we obtained a series of Wor1 interacting *** these proteins, Fun30 could interact with Wor1 both in yeast two hybrid systemand in C. albicans in vivo system. The Fun30 in S. cerevisia is a chromatin remodeler,but the function in C. albicans is unknown. We want to investigate that how Fun30 is participatedin white-opaque switching and Wor1 regulation network. Deletion of Fun30 impairedopaque cell formation and reduced white to opaque switching frequency and facilitatedopaque cells switching to white cells. Ectopic expression of Fun30 in C. albicans significantlyincreased the white to opaque switching frequency. Furthermore, we found thatFun30 stimulating opaque cell phenotype could not bypass the requirement of Wor1 andFun30 could bind at Wor1 promoter region in opaque cells. The virulence assay showed that fun30Δ is a lower toxicity in mouse model. Our results demonstrated that Fun30 plays important roles in white-opaque switching.

关键词： Fun30 Wor1

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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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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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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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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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MicroRNA-210 negative regulates Notch signaling mediatedinflammatory response in murine bone marrow derived macrophage

MicroRNA-210 negative regulates Notch signaling mediatedinfl...

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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 Huang Junlong Zhao Hongyan Qin Hua Han Department of Medical Genetics and Developmental Biology Fourth Military Medical University

macrophage polarizationMacrophages participate in innate immune responses through different activation patterns,resulting in inflammatory reactions, tissue repairs, and other different biological *** preliminary studies using tumor models, we found that activating Notch signalingpromotes M1 polarization, while blocking Notch signaling promotes M2 polarization, suggestingthat Notch signaling is an important factor in the regulation of macrophage ***(miRNAs) are highly conversed, small non-coding RNA molecules thatplay an essential role in the regulation of gene expression and functions. By microarrayanalysis between the macrophages obtained from RBP-J, the key transcription factor ofNotch signaling, knockout mice and normal mice, we found that there were a set of differentiallyexpressed miRNAs, which suggested that the regulation of macrophage activationby Notch signaling may work through miRNA. In our present study, we found that miR-210, a Notch positive-regulated miRNA in murine bone marrow derived macrophage underinflammatory stimuli, worked as a negative regulator in inflammatory response mediatedby Notch *** real-time quantitative PCR, we found that miR-210 increased in M1 polarization inducedby LPS and IFN-r, but significantly downregulated in Lyz2-Cre-Rbpj-/- *** miR-210 mimic transfection, LPS/IFN-r-induced expression of proinflammatory cytokines,such as TNF-a, IL-6 and iNOS, were significantly reduced, which were significantlyincreased after miR-210 inhibitor transfection. Meanwhile, Bacterial phagocytic and killingexperiments showed that after miR-210 mimic transfection, the ability of macrophae uponphagocytic and bacterial killing were significantly decreased. Mixed lymphocyte reactionshowed that antigen-presenting ability of macrophages were significantly reduced afteroverexpression of miR-210. Assay of luciferase reporter gene expression demonstratedthat transfection of miR-210 mimics significantly inhibited

关键词： MicroRNA-210 Notch

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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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