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Crystal structure of Rot, a global regulator of virulence genes in Staphylococcus aureus

Crystal structure of Rot, a global regulator of virulence ge...

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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：朱玉威范晓娇滕脉坤李旭中国科学技术大学

Staphylococcus aureus is a highly versatile pathogen that can infect the tissue ofhuman by producing a large arsenal of virulence factors,which was tightly regulated by acomplex regulatory ***,sharing sequence similarity to SarA homologues,is aglobal regulator that regulates numerous virulence ***,the recognition modelof Rot to the promoter region of target genes and the putative regulation mechanism *** this study,we report the 1.77resolution X-ray crystal structure of *** structure reveals that two Rot molecules form a compact homodimer,each of whichcontains a typical helix-turn-helix module and aβ-hairpin motif connected by a *** fluorescence polarization results indicate that Rot preferentially recognizes ATrichdsDNA with≈30 base pair nucleotides,and the conserved positively charged residueson the winged-helix motif are vital for binding to the AT-rich *** propose that theDNA recognization model of Rot may be similar to that of SarA,SarR and SarS,in whichthe helix-turn-helix motifs of each monomer interact with the major grooves of targetdsDNA and the winged motifs contact the minor ***,the structureshows that Rot adopts a novel dimerization model differing from that of other SarA *** expected,perturbation of the dimer interface abolishes the dsDNA binding abilityof Rot,suggesting that Rot functions as a *** addition,we further confirm our resultsin vivo by measuring the transcriptional regulation ofα-toxin,a major virulence factorsproduced by most *** strains.

关键词： Rot virulence genes Staphylococcus aureus

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MIF intersubunit disulfide mutant antagonist supports activation of CD74 by endogenous MIF trimer at physiologic concentrations

MIF intersubunit disulfide mutant antagonist supports activa...

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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：范成鹏 Elias Lolis 武汉大学 Yale University

Macrophage migration inhibitory factor(MIF)is a proinflammatory *** addition toits known receptor-mediated biological activities,MIF possesses a catalytic site of unknownfunction between subunits of a *** subunit contributes threeβ-strands toadjacent subunits to form a core seven-strandedβ-sheet for each ***,dimers,or trimers have been reported,but the active form that binds and activatesthe MIF receptor(CD74)is still a matter of debate.A cysteine mutant(N110C)thatcovalently locks MIF into a trimer by forming a disulfide with Cys-80of an adjacent subunit is used to study this *** catalytic activity and receptorbinding to CD74 are retained by N110C(locked trimer),but there is no cellular ***-type MIF-induced cellular signaling,in vivo lung neutrophil accumulation,and alveolarpermeability are inhibited with a fivefold excess of *** size-exclusion chromatography with light scattering reveal that N110C can form ahigher-order oligomer in equilibrium with a single locked *** X-ray structure confirmsa local conformational change that disrupts the subunit interface and results in globalchanges responsible for the oligomeric *** structure also confirms these changesare consistent for the partial catalytic and receptor binding *** absence of anypotential monomer and the retention of partial catalytic and receptor binding activitiesdespite changes in conformation(and dynamics)in the mutant support an endogenousMIF trimer that binds and activatesCD74 at nanomolar *** conclusion has implications for therapeutic development.

关键词： MIF strcuture mechanism

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IQGAP1 is involved in cell migration by interacting with γ-cateninin hepatocellular carcinoma

IQGAP1 is involved in cell migration by interacting with γ-...

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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： Xuewen Jin Yuling Liu Weiliang Lu Bing Chen Jingjing Liu Peng Zhang Shufang Liang State Key Laboratory of Biotherapy/Collaborative Innovation Center for Biotherapy West China Hospital Sichuan University West China Hospital Sichuan University

The IQ-domain GTPase-activating protein 1(IQGAP1) is a multifunctional scaffold protein,which interacts with diverse proteins to regulate cell adhesion and cell migration. And theabnormal expression of IQGAP1 widely exists in many cancers, the molecular mechanismof IQGAPs in tumorigenesis remains to clarify. We have found that IQGAP1 is overexpressedin hepatocellular carcinoma(HCC) cell lines and HCC tumor tissues. The expressionlevels of IQGAP1 are positively associated with the metastasis ability of HCC cells. Theup-regulation of IQGAP1 improves the expression of β-catenin and the migration ability ofHepG2 cells. Meanwhile, the knockdown of IQGAP1 by siRNA results in the decrease of β-catenin expression and cell migration in vitro. Furthermore, the protein expression ofIQGAP1 and β-catenin are respectively upregulated in 54.55% and 45.45% of human HCCtumor samples(n=11) compared with the para-cancerous liver tissues. Generally the higher expressions and interactions of IQGAP1 and β-catenin contribute to HCC tumori genesisby promoting cell growth and cell migration.

关键词： IQGAP1 β-catenin Hepatocellular carcinoma Cell migration

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Roles of scaffolding protein Gab1 in regulation of myeloid dendritic cell-mediated allergic asthma

Roles of scaffolding protein Gab1 in regulation of myeloid d...

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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 Yun Xu Dong Cen Jiaqi Xu Xue Zhang Yuehai Ke zhejang University Zhejang University

Scaffolding proteins Gab1 is ubiquitously expressed and belongs to the GAB family(GABs),which integrates multiple signal *** protein family has recently attracted interestthat genetic variants of human GAB1 are considered a risk factor for *** previousresearch has revealed a novel function of Gab2 in mediating mucin gene expression andGoblet cell hyperplasia(GCH),which is an important process in ***,the functionof Gab1 in asthma remains *** asthma exacerbated patients,we found a prominentelevated level of Gab1 in peripheral blood monouclear cells compared to that of ***,we generated conditional Gab1 deletion mice in myeloid cells(Gab1 KO mice)to determinate physiological the role of Gab1 in *** histologyand mucus metaplasia indicated that disruption of Gab1 in dendritic cells(DCs)leads to alleviatedasthmatic *** investigation showed that loss of Gab1results inimpaired DC migration to lymph node,while has no influences on DC maturation、antigenuptake and ***,Gab1 is essentially involved in CCL19-mediatedAKT ***,these finding has characterized a novel of Gab1 in mediating DCmigration and that deregulation of Gab1 signal in lungs predisposes asthmatic inflammation.

关键词： Gab1 dendritic cell allergic asthma

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The M-T Hook Structure Dramatically Increases the Potency of HIV-1 Fusion Inhibitors to Overcome Drug-resistance

The M-T Hook Structure Dramatically Increases the Potency of...

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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：姚雪崔胜医科院病原所

Peptides derived from the C-terminal heptad repeat region(CHR) of HIV-1 gp41 are potentfusion inhibitors by targeting the N-terminal heptad repeat region(NHR). We have recentlyidentified that the unique M-T hook structure preceding the pocketbinding motifof CHR peptides can greatly enhance the binding of inhibitors to the NHR pocket thus improvingthe anti-HIV activity. Here, we incorporated the M-T hook to the N-terminus of Sifuvirtide(SFT), a potent CHR-derived inhibitor currently under phase III clinical trials inChina. The high-resolution crystal structure of the resulting peptide MT-SFT confirmed ahighly conserved M-T hook conformation. Compared to SFT, MT-SFT exhibited significantlyimproved activities to bind to the NHR and to block the formation of six-helix bundle(6-HB) and to inhibit HIV-1 Env-mediated cell fusion, viral entry and replication. Importantly,MT-SFT inhibited SFT- and Enfuvirtide(T20)-resistant HIV-1 variants more efficientlyand had a dramatically increased genetic barrier to induce drug resistance. Therefore, ourstudies have verified that the M-T hook structure offers a general strategy for designingnovel HIV-1 fusion inhibitors and provide new insights into viral entry and inhibition.

关键词： HIV fusion inhibitor

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microRNA expression profiling of the normal and regenerating liver of Chiloscyllium plagiosum

microRNA expression profiling of the normal and regenerating...

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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： conger lu xinyi shi weidong wang zuoming nie jian chen wutong wu zhengbing lv Zhejiang Sci-Tech University

C. plagiosum is an abundant fish that is representative of the cartilaginous class and foundin the southeast coastal region of China. It has a large liver containing many bioactivesubstances. This study aimed to improve our understanding of the mechanisms ofliver regeneration in sharks and to identify the microRNAs that participate in liverregeneration and other liverrelated diseases. To this end, normal and regenerating livertissues from C. plagiosum were harvested 0, 3, 6, 12 and 24 h after partial hepatectomy(PH) and were sequenced using the Illumina/Solexa platform. In total, 309 known microRNAsand 590 novel microRNAs were identified in C. plagiosum. There were 368 microRNAsdifferentially expressed between the normal and regenerating livers. Using targetprediction and GO analysis, most of the differentially expressed microRNAs were assignedto functional categories that may be involved in regulating liver regeneration, suchas cell proliferation, differentiation and apoptosis. Additionally, this study adds severalnovel microRNAs to the database, which will help identify microRNAs in other genetically related species and provides a starting point for future studies aimed at understanding the roles of microRNAs in liver regeneration and other liver diseases.

关键词： deep sequencing differentially expressed microRNA C.plagiosum liver regeneration

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AGEs derived from ribosylation triggering cognitive impairment inrat

AGEs derived from ribosylation triggering cognitive impairme...

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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：吴蓓蓓魏艳赫荣乔 State Key Laboratory of Brain and Cognitive Sciences Institute of Biophysics Chinese Academy of Sciences

Advanced glycation end products(AGEs), products of the irreversible reaction of carbohydratesand protein in the Maillard reaction, accumulate in plasma and tissues in diabetesand renal failure. Evidences also indicate a moderately increased risk of development ofneurodegenerative diseases in diabetic patients. Recent studies showed that, D-Ribose, animportant reducing monosaccharide, is highly active in the glycation of proteins. DribosylatedBSA protein can decrease the cell viability, induce the increasement of ROSlever and apoptosis in the neurotypic cell line SH-SY5 Y. Here, we treated rats with Dribose-glycated products by tail vein injection to explore the relationship between AGEsand cognitive impairment. SD rats injected with D-ribose-glycated BSA for 30 days showhigh blood levels of AGEs. Administration of D-ribose-glycated BSA accelerates AGEsformation in rat brains and induces impairment of spatial learning and memory ability accordingto the performance in Morris water maze test. Furthermore, hyperphosphorylatedTau(pS396, AT8) was observed in brain of rat. Results from immunohistochemistry showthat there were hyperphosphorylated Tau in rat brain. These datas demonstrate that Driboseglycated products are closely related with cognitive impairment. Moreover, the ratsinjected with D-ribose-glycated BSA express anxiety and show different levels of oxidativestress in different tissues. We will continue to investigate the related molecular mechanismin the future.

关键词： AGEs cognitive impairment anxiety oxidative stress

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H19 is involved in regulating VSMC phenotypic switching and proliferation

H19 is involved in regulating VSMC phenotypic switching and ...

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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：孙绍光许馨苗穗兵李芳芳韩梅河北医科大学

H19 is a paternally imprinted gene whose expression produces a .4 kb long noncodingRNA, being one of the most highly abundant and conserved transcripts in mammalian development.H19 is reexpressed in human atherosclerotic plaques, yet its function remainsunknown. Here we show that H19 highly expressed in the rat carotid arteries at 28 daysafter balloon injury by transcriptome sequencing and qRT-PCR analysis, compared with innormal rat carotid arteries. Consistent with in vivo assay, H19 expression level upregulatedin cultured dedifferentiational VSMCs compared with in cultured differentiational VSMCs invitro identified by qRT-PCR. Knockdown of H19 by siRNAs in dedifferentiational VSMCsresulted in their reduced proliferation, and the SM22α（a VSMC differentiational markergene） expression level upregulation detected by Western Blot. In contrast, overexpressionof H19 in differentiational VSMCs resulted in their promoted proliferation, and the SM22αexpression level downregulation. These results suggest that H19 may play an importantrole in the regulation of proliferation and phenotypic switching of VSMCs, which are associated with neointimal formation.

关键词： H19 long noncoding RNA vascular smooth muscle cell phenotypic switchingn eointimal formation

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A systematic genetic polymorphism analysis of the CYP2D6 gene in the Chinese Uighur minority population

A systematic genetic polymorphism analysis of the CYP2D6 gen...

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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：孙文欢王颖莉李树春中央民族大学

Objective: The purpose of this study was to determine the genetic polymorphisms of theCYP2D6 gene and to elucidate the allele distribution pattern in the Chinese Uighur minoritypopulation. Methods: We have selected unrelated healthy minority's adults and establisheda genomic DNA library of Chinese ethnic minorities. We used PCR and bidirectionalsequencing methods to analyze all nine exons of the CYP2D6 gene in 131 Chinese Hansubjects and 76 Uighur subjects from the established library of DNA samples. Results: Intotal, 14 nonsynonymous mutated sites were detected in 207 participants, of which 4 siteswere reported for the first time. We got the gene frequency distribution and genotype frequencydistribution. At the same time, linkage disequilibrium analysis and tagSNP selectionwere performed separately. By using these methods, distinct differences were foundbetween the two ***: This study provides the most comprehensivedata concerning CYP2D6 polymorphisms in the Chinese Uighur minority population todate and increases the number of known alleles;these findings may greatly contribute tothe development of personalized medicine for the Chinese Uighur minority population.

关键词： Chinese Uighur minority population CYP2D6 polymorphism

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Polymorphisms of EPAS1 Gene Associations with High Altitude Polycythemia in Tibetans at the Qinghai-Tibetan Plateau

Polymorphisms of EPAS1 Gene Associations with High Altitude ...

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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：胥瑾 Research Center for High Altitude Medicine Qinghai University Medical School

Objective To test the hypothesis that the polymorphisms in EPAS1 gene are associatedwith the susceptibility to high altitude polycythemia(HAPC) in Tibetans at the Qinghai-Tibetan Plateau. Methods This study enrolled 63 Tibetan HAPE patients,matched 131 healthy Tibetans as control, both of them were from Yushu area in Qinghai where the altitudeis over 3,500 m. Eight SNPs of the EPAS1 gene, including rs12619696, rs13420857,rs2881504, rs4953388, rs13419896, rs4953354, rs10187368 and rs7587138 were genotypedby the Sequenom MassARRAY SNP assays. Results The frequencies of G allele of EPAS1 SNP rs13419896 were significantly higher in HAPC group than in control group(p<0.05),Moreover, we found the A allele of rs12619696 and rs4953354 much more prevalent inHAPC group, and their counterpart homozygotes prevalent in normal Tibetan group(p<0.05).Conclusions The EPAS1 SNP rs13419896 polymorphism G and A allele ofrs12619696 and rs4953354 are associated with susceptibility to HAPC in Tibetans.

关键词： EPAS1 High Altitude Polycythemia

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