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A Viral Protein Suppresses siRNA-directed Interference in Tobacco Mosaic Virus Infection

Acta Biochimica et Biophysica Sinica 2005年第4期37卷 248-253页

作者： Ming-Min ZHAO De-Rong AN Guang-Hua HUANG~1 Zu-Hua HE~2 Jiang-Ye CHEN~1 College of Plant Protection,North West Science and Technology University of Agriculture and Forestry,Shaanxi 712100,china ~1state key laboratory of molecular biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences. chinese academy of sciences.shanghai 200031,china ~2state key laboratory of Plant molecular Genetics.institute of Plant Physiology and Ecology,shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200032.china College of Plant Protection North West Science and Technology University of Agriculture and ForestryShaanxi 712100 China state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China state key laboratory of Plant molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

Plant viruses encode suppressors of post-transcriptional gene silencing (PTGS), an adaptivedefense response that limits virus replication and its spread in *** helper component proteinase (HC-Pro) of the potato virus A (PVA,genus Potyvirus) suppresses PTGS of silenced ***,the effectof HC-Pro on siRNA-directed interference in the tobacco mosaic virus (TMV) was examined by using atransient Agrobacterium tumefaciens-based delivery system in intact *** was shown that the interferenceeffect was completely blocked by co-infiltration with HC-Pro plus siRNA constructs in both systemic andhypersensitive *** the system host,all plants agro-infiltrated with HC-Pro plus siRNA constructsdisplayed the same symptoms as the negative ***,TMV RNA accumulation was found to beabundant in the upper leaves using reverse transcriptase-PCR (RT-PCR) and Northern blot *** thecontrary,plants agro-infiltrated with the siRNA construct alone were free of ***,ourstudy suggests that the transient expression of HC-Pro inhibited the siRNA-directed host defenses againstTMV infection.

关键词： siRNA HC-Pro tobacco mosaic virus suppressor

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Tributyrin inhibits human gastric cancer SGC-7901 cell growth by inducing apoptosis and DNA synthesis arrest

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World Journal of Gastroenterology 2003年第4期9卷 660-664页

作者： Jun Yan Yong-Hua Xu Lab of molecular and cellular Oncology and Lab of molecular cell biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200031,china Lab of molecular and cellular Oncology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China.

AIM: To evaluate the effects of tributyrin, a pro-drug of natural butyrate and a neutral short-chain fatty acid triglyceride, on the growth inhibition of human gastric cancer SGC-7901 ***: Human gastric cancer SGC-7901 cells were exposed to tributyrin at 0.5, 1, 2, 5, 10 and 50 mmol·L-^1 for 24-72h. MTT assay was applied to detect the cell proliferation.[^3H]-TdR uptake was measured to determine DNA *** morphology was observed by electron microscopy and Hoechst-33258 staining. Flow cytometry and terminal deoxynudeotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay were performed to detect tributyrin-triggered apoptosis. The expressions of PARP, Bcl-2 and Bax were examined by Western blot ***: Tributyrin could initiate growth inhibition of SGC-7901 cell in a dose-and time-dependent manner. [^3H]-TdR uptake by SGC-7901 cells was reduced to 33.6% after 48h treatment with 2mmol·L^-1 tributyrin, compared with the control (Pcells in dose-dependent manner. After 48 hours incubation with tributyrin at 2mmol·L^-1, the level of Bcl-2 protein was lowered, and the level of Bax protein was increased in SGC-7901, accompanied by PARP ***: Tributyrin could inhibit the growth of gastric cancer cells effectively in vitro by inhibiting DNA synthesis and inducing apoptosis, which was associated with the downregulated Bcl-2 expression and the up-regulated Bax expression. Therefore, tribublrin might be a promising chemopreventive and chemotherapeutic agent against human gastric carcinogenesis.

关键词：丁酸甘油酯胃癌细胞凋亡 DNA合成抑制 SGC-7901细胞系化学疗法

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Overexpression of a novel gene , Cms1, can rescue the growth arrest of a Saccharomyces cerevisiae mcm10 suppressor

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cell Research 2001年第4期11卷 285-291页

作者： WANG JI WU, JIA RUI WU state key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai Research Center of Life Science, shanghai institutes for biological sciences. chinese academy of sciences. 320 Yue- Yang Road, shanghai 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

MCM10 protein is an essential replication factor involved in the initiation of DNA replication. A mcm10 mutan (mcm10-1) of budding yeast shows a growth arrest at 37 C. In the present work, we have isolated a mcm10-1 suppressor strain, which grows at 37 C. Interestingly, this mcm10-1 suppressor undergoes cell Cycle arrest at 14 C. A novel gene, YLR003c, is identified by high-copy complementation of this suppressor. We called it as Cms1 (Complementation of Mcm 10 Suppressor). Furthermore, the experiments of trans- formation show that cells of mcm10-1 suppressor with high-copy plasmid but not low-copy plasmid grow at 14 C, indicating that overexpression of Cms1 can rescue the growth arrest of this mcm10 suppressor at non-permissive temperature. These results suggest that CMS1 protein may functionally interact with MCM10 protein and play a role in the regulation of DNA replication and cell cycle control.

关键词： Saccharomyces cerevisiae DNA replication Mcm10 Cms1 cell cycle.

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Bcl-2 over-expression and activation of protein kinase C suppress the Trail-induced apoptosis in Jurkat T cells

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cell Research 2001年第2期11卷 101-106页

作者： GUO BEN CHANG, YONG HUA XU institute of biochemistry and cell biology, shanghai institutes for biological sciences. chinese academy of sciences. 320 Yue-yang Road, shanghai 200031, china 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-yang Road Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-yang Road Shanghai 200031 China

Trail, a tumor necrosis factor-related apoptosis-inducing ligand, is a novel potent endogenous activator of the cell death pathway through the activation of cell surface death receptors Trail-R1 and Trail-R2. Its role, like FasL in activation-induced cell death (AICD), has been demonstrated in immune system. However the mechanism of Trail induced apoptosis remains nuclear. In this report, the recombinant Trail protein was expressed and purified. The apoptosis-inducing activity and the regulation mechanism of recombinant Trail on Jurkat T cells were explored in vitro. Trypan blue exclusion assay demonstrated that the recombinant Trail protein actively killed Jurkat T cells in a dose-dependent manner. Trail-induced apoptosis in Jurkat T cells were remarkably reduced by Bcl-2 over expression in BcL-2 gene transfected cells. Treatment with PMA (phorbol 12-myristate 13-acetate), a PKC activator, suppressed nail-induced apoptosis in Jurkat T cells. The inhibition of apoptosis by PMA was abolished by pretreatment with Bis, a PKC inhibitor. Taken together, it was suggested that Bcl-2 over-expression and PMA activated PKC actively down-regulated the Trail-mediated apoptosis in Jurkat T cell.

关键词： trail Jurkat T cells apoptosis Bcl-2 PEA PKC Bis.

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Positive Charges on Lysine Residues of the Extrinsic 18 kDa Protein Are Important to Its Electrostatic Interaction with Spinach Photosystem Ⅱ Membranes

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 737-742页

作者： Jin-Peng GAO Zhen-Hua YONG Feng ZHANG Kang-Cheng RUAN Chun-He XU Gen-Yun CHEN institute of Plant Physiology & Ecology.shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200032,china laboratory of Proteomics,institute of biochemistry and cell biology,shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200031,china College of sciences.Zhejiang University.Hangzhou 310029,china Graduate School of the chinese academy of sciences.shanghai 200032,china institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China Graduate School of the chinese academy of sciences.hanghai 200032 China College of sciences Zhejiang UniversityHangzhou 310029 China laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China

To determine the contribution of charged amino acids to binding with the photosystem Ⅱcomplex(PSⅡ),the amino or carboxyl groups of the extrinsic 18 kDa protein were modified with N-succinimidyl propionate(NSP)or glycine methyl ester(GME)in the presence of a water-soluble carbodiimide,*** on isoelectric point shift,4-10 and 10-14 amino groups were modified in the presence of2 and 4 mM NSP,***,3-4 carboxyl groups were modified by reaction with *** of negatively charged carboxyl groups with GME did not alter the binding activity of theextrinsic 18 kDa ***,the NSP-modified 18 kDa protein,in which the positively charged aminogroups had been modified to uncharged methyl esters,failed to bind with the PSⅡ membrane in the presenceof the extrinsic 23 kDa *** defect can not be attributed to structural or conformational alterationsimposed by chemical modification,as the fluorescence and circular dichroism spectra among native,GME-and NSP-modified extrinsic 18 kDa proteins were ***,we have concluded that the positive chargesof lysyl residues in the extrinsic 18 kDa protein are important for its interaction with PSⅡ membranes in thepresence of the extrinsic 23 kDa ***,it was found that the negative charges of carboxylgroups of this protein did not participate in binding with the extrinsic 23 kDa protein associated with PSⅡmembranes.

关键词： extrinsic 18 kDa protein circular dichroism electrostatic interaction chemical modification

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Searching for Potential Drug Targets in Two-component and Phosphorelay Signal-transduction Systems using Three-dimensional Cluster Analysis

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Acta Biochimica et Biophysica Sinica 2005年第5期37卷 293-302页

作者： Xiao-Hui CAI Qing ZHANG Shuo-Yong SHI Da-Fu DING key laboratory of Proteonmics,institute of biochemistry and cell biology,Stanghai institutes for biological sciences. chinese academy of sciences.Graduate School of the chinese academy of sciences. shanghai 200031,china key laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Two-component and phosphorelay signal transduction systems are central components in thevirulence and antimicrobial resistance responses of a number of bacterial and fungal pathogens;in somecases,these systems are essential for bacterial growth and ***,we analyze in detail the conservedsurface residue clusters in the phosphotransferase domain of histidine kinases and the regulatory domain ofresponse regulators by using complex structure-based three-dimensional cluster *** also investigatethe protein-protein interactions that these residue clusters participate *** Spo0B-Spo0F complex structurewas used as the reference structure,and the multiple aligned sequences of phosphotransferases and responseregulators were paired *** results show that a contiguous conserved residue cluster isformed around the active site,which crosses the interface of histidine kinases and response *** residue clusters of phosphotransferase and the regulatory domains are directly involved in thefunctional implementation of two-component signal transduction systems and are good targets for thedevelopment of novel antimicrobial agents.

关键词： two-component system phosphorelay histidine kinase response regulator three-dimensional cluster analysis

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Expression of Overlapping PreS1 Fragment Recombinant Proteins for the Determination of Immunogenic Domains in HBsAg PreS1 Region

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 397-404页

作者： Wei-Guo HU, Jun WEI&, Xin-Xiu YANG, Heng-Chuan XIA, Feng LI, and Zu-Chuan ZHANG* key laboratory of Proteomics, institute of biochemistry and cell biology, shanghai institutes for biological sciences.the chinese academy of sciences. shanghai 200031, china key laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of SciencesShanghai 200031 China

In this study, eight preS1 fragments overlapped in preS1 (21–119) region of HBV adr subtype, i.e. preS1 (21–47), preS1 (34–59), preS1 (48–70), preS1 (60–85), preS1 (71–94), preS1 (86–109), preS1 (95–119) and preS1 (21–119), were cloned by PCR, and expressed as GST fusion proteins. These GST- preS1 fusion proteins were highly expressed in soluble form in E. coli, and about 50 to 90 mg soluble fusion proteins were purified from 1 L culture. Using these fusion proteins, the immunogenic domains in preS1 (21– 119) region were identified by Western blot analysis and competitive ELISA. The results showed that the immunogenic domains mainly existed in preS1 (21–59) in N-terminus and preS1 (95–109) in C-terminus, and more importantly, a major immunogenic domain preS1 (34–59), which has much stronger immunogenicity, was identified. It was also supported by the predictions of secondary structure and immunological property in the preS1 (21–119) region. The results here would be helpful for the design of new vaccines against HBV.

关键词： HBV preS1 overlapping fragment immunogenic domain

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Synapse formation and remodeling

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Science china(Life sciences. 2010年第3期53卷 315-321页

作者： LUO ZhenGe institute of Neuroscience and state key laboratory of Neuroscience,shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200031,china institute of Neuroscience and state key laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences

Synapses are specialized structures that mediate information flow between neurons and target cells,and thus are the basis for neuronal system to execute various functions,including learning and *** are around 1011 neurons in the human brain,with each neuron receiving thousands of synaptic inputs,either excitatory or inhibitory.A synapse is an asymmetric structure that is composed of pre-synaptic axon terminals,synaptic cleft,and postsynaptic *** formation involves a number of cell adhesion molecules,extracellular factors,and intracellular signaling or structural *** the establishment of synaptic connections,synapses undergo structural or functional changes,known as synaptic plasticity which is believed to be regulated by neuronal activity and a variety of secreted *** review summarizes recent progress in the field of synapse development,with particular emphasis on the work carried out in china during the past 10 years(1999-2009).

关键词： synapse development neuronal activity synaptic plasticity

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Preparation and Preliminary Application of Monoclonal Antibodies against Trichokirin-S1,a Small Ribosome-inactivating Peptide from the Seeds of Trichosanthes kirilowii

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Acta Biochimica et Biophysica Sinica 2005年第7期37卷 447-452页

作者： Xin-Xiu YANG Feng LI Wei-Guo HU Heng-Chuan XIA Zu-Chuan ZHANG key laboratory of Proteomics.institute of biochemistry and cell biology,shanghai institutes for biological sciences.chinese academy of sciences.shanghai 200031,china key laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Trichokirin-S1,a small ribosome-inactivating peptide recently purified from the seeds ofTrichosanthes kirilowii,has potential clinical applications because of its small molecular *** stablestrains of hybridomas (1F11 and 2A5) that can secrete highly specific monoclonal antibodies (mAbs) againstTrichokirin-S1 have been developed using the hybridoma *** isotypes of these two mAbs,1F11and 2A5,were determined to be IgGand IgG,*** affinity constants,which were measuredby non-competitive ELISA,were found to be 2.3×10~8 Mand 2.8×10~8 M,*** immunoaffinitymethod using 2A5-coupled Sepharose 4B was successfully developed to purify *** twoantibodies have also been used to detect Trichokirin-S1 in Western blot.

关键词： Trichokirin-S1 monoclonal antibody Western blot affinity chromatography

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Progress in glial cell studies in some laboratories in china

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Science china(Life sciences. 2010年第3期53卷 330-337页

作者： DUAN ShuMin institute of Neuroscience,state key laboratory of Neuroscience,shanghai institutes for biological sciences.chinese academy of sciences.shanghai china,200031 institute of Neuroscience State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences

Glial cells in the central nervous system(CNS) consist of a heterogeneous population of cell types,each characterized by distinct morphological features,physiological properties,and specific *** contrast to the previous view that glial cells were passive elements in the brain,accumulating evidence suggests that glial cells are active participants in various brain functions and brain *** review summarizes recent progress of glial cell studies from several groups in china,ranging from studies about the mechanisms of neuron-glia crosstalking to investigations on the roles of glial cells in various CNS disorders.

关键词： neuron-glia interaction ATP plasticity pain ischemia neural injury and repair

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