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Cell Research 2001年第1期11卷 61-67页

作者： MA YINGHUA, JIA HUA HU, WEN JUAN ZHAO, JIAN FEI, yuN yu, XIAO GANG ZHOU, ZHEN TONG MEI, li HE GUO (institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china) (shanghai Inst institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China shanghai institute of physiology Chinese Academy of Sciences Shanghai China

γ-aminobutyric acid (GABA) is the principal inhibitory neurotransmitter, and the GABAergic synaptic transmission is normally terminated by the rapid uptake through GABA transporters. With transgenic mice ubiquitously overexpressing GABA transporter subtype I (GAT1), the present study explored the pathophysiological role of GAT1 in epileptogenesis. Though displaying no spontaneous seizure activity, these mice exhibit altered electroencephalographic patterns and increased susceptibility to seizure induced by kainic acid. In addition, the GABAA receptor and glutamate transporters are up-regulated in transgenic mice, which perhaps reflects a compensatory or corrective change to the elevated level of GAT1. These preliminary findings support the hypothesis that excitatory and inhibitory neurotransmission, and seizure susceptibility can be altered by neurotransmitter transporters.

关键词： γ-aminobutyric acid transporter seizure susceptibility kainic acid electroencephalography transgenic mice

Six Specific Lysihe Residues are Crucial in Maintaining the Structure and Function of Soluble Manganese Stabilizing Protein

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Acta Biochimica et Biophysica Sinica 2006年第9期 611-619页

作者： Jin-Peng GAO~(1,2#) feng ZHANG~(1,3#) li ZHANG~4 Yin-Long GUO~4 Kang-Cheng RUAN~5 De-An JIANG~3* Chun-He XU~1 1 institute of Plant physiology and Ecology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200032,china 2 Graduate School of the chinese academy of sciences,shanghai 200032,china 3 College of life sciences,Zhejiang University,Hangzhou 310029,china 4 shanghai Mass Spectrometry Center,shanghai institute of Organic Chemistry,chinese academy of sciences,shanghai 200031,china 5 Laboratory of Proteomics,institute of Biochemistry and Cell Biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china

When manganese stabilizing protein （MSP） was treated with 0.5mM N-succinimidyl propionate（NSP）,the rebinding ability and oxygen-releasing capabilities of the modified MSP were not altered,in spiteof changes of MSP surface Lys ***,far-ultraviolet circular dichroism and intrinsicfluorescence spectra analysis revealed that 0.5 mM NSP-modified MSP retained most of its native secondaryand tertiary *** of the sites of NSP modification by Staphylococcus V8protease digestion ofthe modified protein,as well as analysis by matrix-assisted laser desorption ionization-time of flight massspectrometry,indicated that seven Lys residues were *** results suggested that these residues arenot absolutely essential to the structure and function of ***,when the NSP concentration wasincreased to 4mM,the modified MSP was unable to bind photosystem Ⅱ and completely lost its *** far-ultraviolet circular dichroism and intrinsic fluorescence spectra analysis revealed a clearconformational change in MSP after 4 mM NSP treatment,suggesting that some Lys residues are involved inmaintaining the structure and function of *** by matrix-assisted laser desorption ionization-timeof flight mass spectrometry indicated that another six Lys residues,namely Lys20,Lys101,Lys196,Lys207,Lys130 （or Lys137） and Lys66 （or Lys76）,were modified by 4 mM ***,these six Lys residuesare crucial in maintaining the structure and function of soluble MSP.

关键词： chemical modification fluorescence spectrum circular dichroism spectrum manganese stabilizing protein Lys residue

Transgenic mice overexpressingγ-aminobutyric acid transporter subtypeⅠ develop obesity

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Cell Research 2000年第4期10卷 303-310页

作者： MA YING HUA JIA HUA HU XIAO GANG ZHOU RUO WANG ZENG ZHEN TONG MEI JIAN FEI li HE GUO institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of Science, shanghai 200031, china shanghai institute of institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Science Shanghai China shanghai institute of physiology Chinese Academy of Science Shanghai China

Transgenic mice ubiquitously overexpressing murine γ aminobutyric acid transporter subtype Ⅰ were created. Unexpectedly, these mice markedly exhibited heritable obesity, which features significantly increased body weight and fat deposition. Behavioral examination revealed that transgeinc mice have slightly reduced spontaneous locomotive capacity and altered feeding pattern. Tills preliminary finding indicates that the inappropriate level of γ-aminobutyric acid transporters may be directly or indirectly involved in the pathogenic mechanism underlying certain types of obesity.

关键词： γ-aminobutyric acid transporter subtype Ⅰ transgenic mouse obesit€

Role of oxyR from Sinorhizobium meliloti in Regulating the Expression of Catalases

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Acta Biochimica et Biophysica Sinica 2005年第6期37卷 421-428页

作者： li EUO~(1.3) Ming-Sheng QI~(1.3) shi-Yi YAO~2 Hai-Ping CHENG~2 Jia-Bi ZHU~1 Guan-Qiao yu~1 1 National Laboratory of Plant Molecular Genetics, institute of Plant physiology and Ecology, shanghai institutes for biological sciences.chinese academy of sciences, shanghai 200032. china 2 biological sciences Department, Lehman College, the City University of New York, Bronx, NY 10468. USA, 3 Graduate School of the chinese academy of sciences, shanghai 200032. china National Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China Graduate School of the chinese academy of sciencesshanghai 200032 China biological sciences Department Lehman College the City University of New YorkBronx NY 10468 USA

The process of symbiotic nitrogen fixation results in the generation of reactive oxygen speciessuch as the superoxide anion（O2-）and hydrogen peroxide（H2O2）.The response of rhizobia to these toxicoxygen species is an important factor in nodulation and nitrogen *** Sinorhizobium meliloti,oneoxyR homologue and three catalase genes,katA,katB,and katC were detected by sequence *** gene is located next to and divergently from katA on the *** investigate the possible rolesof oxyR in regulating the expression of catalases at the transcriptional level in ***,an insertion mutantof this gene was *** mutant was more sensitive and less adaptive to H2O2than the wild typestrain,and total catalase/peroxidase activity was reduced approximately fourfold with the OxyR mutationrelative to *** activities of KatA and KatB and the expression of katA::lacZ and katB::lacZ promoterfusions were increased in the mutant strain compared with the parental strain grown in the absence of H2O2,indicating that katA and katB are repressed by ***,when exposed to H2O2,katA expressionwas also increased in both *** and Escherichia *** exposed to H2O2,OxyR is convertedfrom a reduced to an oxidized form in *** concluded that the reduced form of OxyR functions as arepressor of katA and katB ***,in the presence of H2O2,reduced OxyR is converted to theoxidized form of OxyR that then results in increased katA *** further showed that oxyR expressionis autoregulated via negative feedback.

关键词： Sinorhizobium meliloti OxyR catalase katA H2O2

The spike protein of severe acute respiratory syndrome (SARS) is cleaved in virus infected Vero-E6 cells

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Cell Research 2004年第5期14卷 400-406页

作者： Xiao Dong WU 1*, Bo SHANG1*, Rui Fu YANG2*, Hao yu3*, Zhi Hai MA1*, Xu SHEN3*, Yong Yong JI1, Ying liN , Ya Di WU1, Guo Mei liN1, lin TIAN1, Xiao Qing GAN1, Sheng YANG2, Wei Hong JIANG2, 1 Er Hei DAI4, Xiao Yi WANG4, Hua liang JIANG3, You Hua XIE1, Xue liang ZHU1, Gang PEI1, lin li1, Jia Rui WU1**, Bing SUN 1** 1institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 yueyang Road, shanghai 200031, china. 2institute of Microbiology and Epidemiology, institute of Basic Medical sciences, academy of Military Medical sciences, Beijing 100071, china. 3institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin Road, shanghai 200031, china. 3institute of Materia Medica, shanghai institutes for biological sciences, chinese academy of sciences, 320 Taiyuan Road, shanghai 200031, china. institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China.

Spike protein is one of the major structural proteins of severe acute respiratory syndrome-coronavirus. It is essential for the interaction of the virons with host cell receptors and subsequent fusion of the viral envelop with host cell membrane to allow infection. Some spike proteins of coronavirus, such as MHV, HCoV-OC43, AIBV and BcoV, are proteolytically cleaved into two subunits, S1 and S2. In contrast, TGV, FIPV and HCoV-229E are not. Many studies have shown that the cleavage of spike protein seriously affects its function. In order to investigate the maturation and proteolytic processing of the S protein of SARS CoV, we generated S1 and S2 subunit specific antibodies (Abs) as well as N, E and 3CL protein-specific Abs. Our results showed that the antibodies could efficiently and specifically bind to their corresponding proteins from *** expressed or lysate of SARS-CoV infected Vero-E6 cells by Western blot analysis. Furthermore, the anti-S1 and S2 Abs were proved to be capable of binding to SARS CoV under electron microscope observation. When S2 Ab was used to perform immune precipitation with lysate of SARS-CoV infected cells, a cleaved S2 fragment was detected with S2-specific mAb by Western blot analysis. The data demonstrated that the cleavage of S protein was observed in the lysate, indicating that proteolytic processing of S protein is present in host cells.

关键词： SARS-CoV spike protein nucleocapsid protein envelope protein 3CL protease polyclonal antibody.

General gambogic acids inhibited growth of human hepatoma SMMC-7721 cells in vitro and in nude mice

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Acta Pharmacologica Sinica 2004年第6期25卷 67-72页

作者： Qing-long GUO2, Qi-dong YOU3, Zhao-qiu WU, Sheng-tao yuAN4,li ZHAO Department of physiology, Department of Medicinal Chemistry, china Pharmaceutical University, Nanjing 210009 3 4institute of Materia Medica, shanghai institutes for biological sciences, chineseacademy ofsciences, shanghai 200031, china Department of physiology China Pharmaceutical University Nanjing 210009 Department of Medicinal Chemistry China Pharmaceutical University Nanjing 210009 institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 上海 200031

AIM: To study the inhibitory effect of general gambogic acids (GGA) on transplantation tumor SMMC-7721 in experimental animal model and SMMC-7721 cells in vitro. METHODS: Anti-tumor activity of GGA in the experi- mental transplantation tumor SMMC-7721 was evaluated by relative tumor growth ratio. Cell morphology was observed with inverted microscope and electron microscope. Cell proliferation was measured by MTT assay and the telomerase activity was determined by PCR. RESULTS: In vivo study indicated that GGA (2, 4, and 8 mg/kg, iv, 3 times per week for 3 weeks) displayed an inhibitory effect on the growth of transplantation tumor SMMC- 7721 in nude mice compared with the normal saline group (Pliferation of SMMC-7721 cells in vitro. GGA 2 mg/L dramatically changed morphology of SMMC-7721 cells and inhibited the telomerase activity in SMMC-7721 cells. CONCLUSION: GGA had inhibitory effect on the growth of SMMC-7721, which might be related to its inhibition of telomerase activity.

关键词： gamboge gambogic acid hepatoma SMMC-7721 cells telomerase nude mice

Preliminary studies on differential defense responses induced during plant communication

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Cell Research 2005年第3期15卷 187-192页

作者： Jin Ying PENG1,3,Zhong Hai li2,Hui XIANG1,3,Jian Hua HUANG1,3,shi Hai JIA1,3,Xue Xia MIAO1,Yong Ping HUANG1* 1institute of Plant physiology and Ecology,shanghai institute for biological sciences,Graduate School of the chinese academy of sciences,chinese academy of sciences,300 fenglin Road,shanghai 200032,china 2chinese National Human Genome Center at shanghai,250 Bibo Road,shanghai 201203,china 3Graduate School of the chinese academy of sciences,Beijing 100039,china institute of Plant physiology and Ecology Shanghai Institute for Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of Sciences Shanghai China Graduate School of the chinese academy of sciences Beijing China chinese National Human Genome Center at shanghai Shanghai China

We compared the expression patterns of three representative genes in undamaged tomato and tobacco plants in response to exposure to either tomato or tobacco fed on by Helicoverpa armigera (cotton bollworm). When tomato and tobacco, two species of one family, were incubated in the chambers with the tomato plants damaged by the cotton bollworm, the expression of the PR1, BGL2, and PAL genes was up-regulated in leaves of both plants. However, the levels of gene expression were significantly higher in the tomato than that in the tobacco. In addition, the activities of enzymes, peroxidase, polyphenol oxidase, and lipoxygenase were found to be higher in the tomato than those in the tobacco. Similar results were obtained when the damaged plants were replaced by the tobacco.

关键词： plant-plant communication tomato (Lycopersicon esculentum) tobacco (Nicotiana tabacum) cotton boll- worm (Helicoverpa armigera) inducible defenses.

Alpha-picolinic acid,a fungal toxin and mammal apoptosis-inducing agent,elicits hypersensitive-like response and enhances disease resistance in rice

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Cell Research 2004年第1期14卷 27-33页

作者： Hai Kuo ZHANG~1, Xin ZHANG~1, Bi Zeng MAO~(2,) Qun li~1, Zu Hua HE~(1,2*) ~1SHARF and National Laboratory of Plant Molecular Genetics, shanghai institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200032, china. ~2Biotechnology institute, Zhejiang University, Hangzhou 310029, china. SHARF and National Laboratory of Plant Molecular Genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Biotechnology institute Zhejiang University Hangzhou China

Alpha-picolinic acid (PA), a metabolite of tryptophan and an inducer of apoptosis in the animal cell, has been reported to be a toxin produced by some of plant fungal pathogens and used in screening for disease resistant mutants. Here, we report that PA is an efficient apoptosis agent triggering cell death of hypersensitive-like response in planta. Confirmed by Fluorescence Activated Cell Sorter (FACS), rice suspension cells and leaves exhibited programmed cell death induced by PA. The PA-induced cell death was associated with the accumulation of reactive oxygen species that could be blocked by diphenylene iodonium chloride, indicating that the generation of reactive oxygen species was NADPH-oxidase dependent. We also demonstrated the induction of rice defense-related genes and subsequent resistant enhancement by PA against the rice blast fungus Magnaporthe grisea. Hence, it was concluded that the PA-stimulated defense response likely involves the onset of the hypersensitive response in rice, which also

关键词： picolinic acid cell death hypersensitive responses Magraporche grisea rice.

Salt-responsive genes in rice revealed by cDNA microarray analysis

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Cell Research 2005年第10期 50-64页

作者： Dai Yin CHAO1,3, Yong Hai LUO1,3, Min shi1, Da LUO1,2, Hong Xuan liN1,2,* 1National Key Laboratory of Plant Molecular Genetics, shanghai institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin Road, shanghai 200032, china, 2SHARF Laboratory, shanghai institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin Road, shanghai 200032, china, 3Graduate School of the chinese academy of sciences (D.C.), 19 yuquan Road, Beijing 100039, china

We used cDNA microarrays containing ~9,000 unigenes to identify 486 salt responsive expressed sequence tags (ESTs) (representing ~450 unigenes) in shoots of the highly salt-tolerant rice variety, Nona Bokra (Oryza sativa L. ssp. Indica pv. Nona). Some of the genes identified in this study had previously been associated with salt stress. However the majority were novel, indicating that there is a great number of genes that are induced by salt exposure. Analysis of the salt stress expression profile data of Nona provided clues regarding some putative cellular and molecular processes that are undertaken by this tolerant rice variety in response to salt stress. Namely, we found that multiple transcription factors were induced during the initial salt response of shoots. Many genes whose encoded proteins are implicated in detoxification, protectant and transport were rapidly induced. Genes supporting photosynthesis were repressed and those supporting carbohydrate metabolism were altered. Commonality among the genes induced by salt exposure with those induced during senescence and biotic stress responses suggests that there are shared signaling pathways among these processes. We further compared the transcriptome changes of the salt-sensitive cultivar, IR28, with that of Nona rice. Many genes that are salt responsive in Nona were found to be differentially regulated in IR28. This study identified a large number of candidate functional genes that appear to be involved in salt tolerance and further examination of these genes may enable the molecular basis of salt tolerance to be elucidated.

关键词： cDNA microarray rice salt stress signaling crosstalk transcriptome.

Cloning and Identification of Methionine Synthase Gene from Pichia pastoris

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Acta Biochimica et Biophysica Sinica 2005年第6期37卷 371-378页

作者： Lan HUANG~1 Dong-Yang li~2 Shao-Xiao WANG~2 shi-Ming ZHANG~2 Jun-Hui CHEN~1 Xiang-Fu WU~2 1 Stute Key Laboratory of Pharmaceutical Biotechnology,Department of Biochemistry.Nanjing University. Nanjing 210093, china 2 institute of Biochemistry and Cell Biology.shanghai institutes.for biological sciences, chinese academy of sciences. shanghai 200031, china ter for Research. Geismger Health System.Danville. PA 17822.USA State Key Laboratory of Pharmaceutical Biotechnology Department of Biochemistry Nanjing University Nanjing 210093 China institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Methionine synthase(MS)is grouped into two *** One MS(MetH)and Class TwoMS(MetE)share no homology and differ in their catalytic *** on the conserved sequences ofmetE genes from different organisms,a segment of the metE gene was first cloned from Pichia pastorisgenomic DNA by PCR,and its 5'and 3'regions were further cloned by 5'-and 3'-rapid amplification ofcDNA ends(RACE),*** assembled sequence reveals an open reading frame encoding apolypeptide of 768 residues,and the deduced product shares 76% identity with MetE of *** methionine synthase(PpMetE)consists of two domains common to *** site is located in the C-terminal domain,in which the residues involved in the interaction of zinc withsubstrates are *** expression of PpMetE in *** was achieved,and the heterolo-gous expression of PpMetE in the *** strain XJB3-1D that is MetE-defective restored the growthof the mutant on methionine-free minimal *** gene sequence has been submitted to GenBank/EMBL/DDBJ under accession ***601648.

关键词： methionine synthase Pichia pastoris rapid amplification of cDNA end(RACE) functional complementation