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Identification of AtENT3 as the main transporter for uridine uptake in Arabidopsis roots

Cell Research 2006年第4期16卷 377-388页

作者： Kun Ling Chen~(1,2,*) Min Xin Xu~(2,3,*) Guang Yong Li~(2,3,*) Hui Liang~2 Zong Liang Xia~(2,3) Xin Liu~2 Ji Shu Zhang~1 Ai Min Zhang~2 Dao Wen Wang~2 ~1school of Life sciences,Northwest Agricultural and Forestry University,Yangling 712.00,china ~2Center for Plant Gene Research(Beijing)and the State Kev Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics and Developmental Biology,chinese academy of sciences,Datun road,Chaovang district,Beijing 100101,china ~3graduate school of the chinese academy of sciences,yuquan road,shijingshan district,Beijing {12.,china school of Life sciences Northwest Agricultural and Forestry University Yangling China Center for Plant Gene Research (Beijing) and the State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Datun Road Chaoyang District Beijing China graduate school of the chinese academy of sciences Yuquan Road Shijingshan District Beijing China

Previous studies have shown that Arabidopsis equilibrative nucleoside transporters （AtENTs） possess transport activitieswhen produced in yeast cells and are differentially expressed in Arabidopsis ***,we report further analysis onthe nucleoside transport activities and transcriptional patterns of AtENT *** recombinant proteins of AtENTs3,6,and 7,but not those of AtENTs 1,2,4,and 8,were found to transport thymidine with high *** to previ-ous suggestion that AtENT1 may not transport uridine,this work showed that recombinant AtENT1 was a pH-dependentand high-affinity transporter of *** grown on MS plates,the AtENT3 knockout plants were more tolerant tothe cytotoxic uridine analog 5-fluorouridine than wild-type plants and the knockout plants ofAtENT 1 or ***-sistent with this observation,the AtENT3 knockout line exhibited a significantly decreased ability to take up [3H]uridinevia the roots when compared with wild-type plants and the plants with mutated AtENT 1 or *** indicates thatAtENT3,but not AtENTs 1 and 6,is the main transporter for uridine uptake in Arabidopsis *** transcription ofAtENTs 1,3,4,6,7,and 8 was regulated in a complex manner during leaf development and *** contrast,thesix AtENT members were coordinately induced during seed *** work provides new information on thetransport properties of recombinant AtENT proteins and new clues for future studies of the in vivo transport activitiesand physiological functions of the different ENT proteins in Arabidopsis plants.

关键词： Arabidopsis equilibrative nucleoside transporter expression pattern nucleoside transport functional analysis

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Expression of Monomeric Insulin Precursor in Pichia pastoris and Its Conversion into Monomeric B27 Lys Destripeptide Insulin by Tryptic Hydrolysis

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 234-240页

作者： Jin-Guo DING~(1,3) Jian FEI~2 Da-Fu CUI~1 You-Shang ZHANG~1 ~1Institute of Biochemistry and Cell Biology,and ~2Model Organism Research Center,Shanghai Institutes for Biologicol sciences, chinese academy of sciences,Shanghai 200031,china ~3graduate school of the chinese academy of sciences,Shanghai 200031,china Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China graduate school of the chinese academy of sciences Shanghai 200031 China Model Organism Research Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from itsprecursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient *** monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium *** by a combination of hydrophobic,size-exclusion,and ion-exchange *** purifiedMIP was converted,by tryptic hydrolysis,to B27 Lys DTrI,which was then purified by ion-exchangechromatography to homogeneity as assessed by native gel electrophoresis,HPLC,amino acid composition,and electrospray mass-spectrometric analysis.B27 Lys DTrI exhibited superior monomeric properties insize-exclusion *** yield of MIP was 200 mg per liter of culture,and the overall yield ofpurified B27 Lys DTrI from the crude MIP was 70%.The in vivo biological activity of B27 Lys DTrI asdetermined by the mouse convulsion assay was 21 U/mg,identical to that obtained by semisynthesis.

关键词： monomeric B27 Lys destripeptide insulin tryptic hydrolysis Pichia pastoris

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Molecular and functional characterization of sulfiredoxin homologs from higher plants

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Cell Research 2006年第3期16卷 287-296页

作者： Xian Peng Liu~(1,2) Xue Ying Liu~(1,2) Juan Zhang~(1,2) Zong Liang Xia~(1,2) Xin Liu~1 Huan Ju Qin~1 Dao Wen Wang~11 The State Key Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics and Developmental Biology,chinese academy of sciences,Datun road,Chaoyang district,Beijing 100101,china 2 graduate University of chinese academy of sciences,yuquan road,shijingshan district,Beijing {12.,china The State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Chaoyang District China graduate University of chinese academy of sciences Shijingshan District China

By reducing cysteine-sulfinic acid in oxidized peroxiredoxin,sulfiredoxin (Srx) plays an important role in oxidationstress resistance in yeast and human ***,we report the first molecular and functional characterization of Srxhomolog from higher *** analysis revealed the presence of potential Srx encoding sequences in bothmonocot and dicot plant *** plant Srx proteins exhibited significant identities to their orthologs from yeastand human,and contained the conserved signature sequence and residues essential for ***,unlike yeastand human orthologs,plant Srxs were all predicted to possess chloroplast transit peptide in their primary *** proteins from Arabidopsis and rice (designated as AtSrx and OsSrx,respectively) complemented functional defi-ciency of Srx in the SRXl deletion yeast cells.A GFP fusion protein of AtSrx was targeted to chloroplast in Arabidopsismesophyll *** transcription occurred in both vegetative and reproductive organs,and the highest transcriptlevel was detected in *** oxidation stress,AtSrx transcript level was substantially increased,which paralleledwith enhanced transcription of 2-Cys peroxiredoxins that have been found essential in maintaining chloroplast *** addition to oxidation stress,osmotic/water deficit or cold treatments also raised AtSrx transcript ***-sistent with above findings,the knock-out mutant of AtSrx was significantly more susceptible to oxidation stress thanwild type Arabidopsis *** together,the results of this work indicate the existence of functional Srx homolog inhigher plants that is essential for plants to cope with oxidation stress.

关键词： Arabidopsis sulfiredoxin chloroplast reactive oxygen species stress

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AtbHLH29 of Arabidopsis thaliana is a functional ortholog of tomato FER involved in controlling iron acquisition in strategy I plants

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Cell Research 2005年第8期15卷 613-621页

作者： You Xi YUAN1,2, Juan ZHANG1,2, Dao Wen WANG1, Hong Qing LING1,* 1The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, chinese academy of sciences, Datun road, Chaoyang district, Beijing 100101, china 2graduate school of the chinese academy of sciences, yuquan road, {12., Beijing 1.0039, china The State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China graduate school of the chinese academy of sciences Beijing China

AtbHLH29 of Arabidopsis, encoding a bHLH protein, reveals a high similarity to the tomato FER which is proposed as a transcriptional regulator involved in controlling the iron deficiency responses and the iron uptake in tomato. For identification of its biological functions, AtbHLH29 was introduced into the genome of the tomato FER mutant T3238fer mediated by Agrobacterium tumefaciencs. Transgenic plants were regenerated and the stable integration of AtbHLH29 into their genomes was confirmed by Southern hybridization. Molecular analysis demonstrated that expression of the exogenous AtbHLH29 of Arabidopsis in roots of the FER mutant T3238fer enabled to complement the defect functions of FER. The transgenic plants regained the ability to activate the whole iron deficiency responses and showed normal growth as the wild type under iron-limiting stress. Our transformation data demonstrate that AtbHLH29 is a functional ortholog of the tomato FER and can completely replace FER in controlling the effective iron acquisition in tomato. Except of iron, FER protein was directly or indirectly involved in manganese homeostasis due to that loss functions of FER in T3238fer resulted in strong reduction of Mn content in leaves and the defect function on Mn accumulation in leaves was complemented by expression of AtbHLH29 in the transgenic plants. Identification of the similar biological functions of FER and AtbHLH29, which isolated from two systematically wide-diverged “strategy I” plants, suggests that FER might be a universal gene presented in all strategy I plants in controlling effective iron acquisition system in roots.

关键词： tomato AtbHLH29 iron uptake Arabidopsis FER plant nutrition.

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Mass Spectrometric Evidence of Heparin Disaccharides for the Catalytic Characterization of A Novel Endolytic Heparinase

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 840-844页

作者： Yapeng CHAO1,3, Shaoxiang XIONG2, Xiulan CHENG1, and Shijun QIAN1* 1Institute of Microbiology, chinese academy of sciences, Beijing 100080, china 2Center of Mass Spectrometry, Institute of Chemistry, chinese academy of sciences, Beijing 100080, china 3graduate school of the chinese academy of sciences, Beijing 1.0039, china Institute of Microbiology Chinese Academy of SciencesBeijing 100080 China graduate school of the chinese academy of sciencesBeijing 1.0039 China Center of Mass Spectrometry Institute of Chemistry Chinese Academy of SciencesBeijing 100080 China

Heparinase from different sources can eliminate heparin or/and heparan sulfate into various low-molecular weight heparins with different characteristics. Porcine intestinal mucosa heparin was de- graded into a series of oligosaccharides by a novel heparinase from the species Sphingobacterium. Disaccha- ride components from the digests were separated and purified by ultrafiltration and HPLC. Five major peaks appeared as three types according to their retention time. The mass spectrometry of peak I mainly gave the non-sulfated disaccharide with the mass of 379 Da. Peak II and III were indicated as two major monosulfated disaccharides with molecular mass of 417 and 459 Da respectively. Moreover, the peak III represented an N- acetyl disaccharide. Both peak IV and V showed the same mass of 496 Da, hinting that they were disulfate- substituted disaccharides. No trisulfate-substituted disaccharides were detected in the mixture of the heparin digest though they were abundant in the heparin structure. The results revealed that the heparinase might specifically cut the sites with low sulfated domain in heparin.

关键词： electrospray ionization-mass spectrometry heparin disaccharides heparinase high perfor- mance liquid chromatography Sphingobacterium sp.

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Transformations and non-degenerate maps

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Science china Mathematics 2005年第Z1期48卷 195-205页

作者： LI Baokui & WANG Yuefei Institute of Mathematics, academy of Mathematics and System sciences, chinese academy of sciences, Beijing 100080, china graduate school, chinese academy of sciences, Beijing 100080, china Institute of Mathematics Academy of Mathematics and System Sciences Chinese Academy of Sciences Beijing China graduate school Chinese Academy of Sciences Beijing China

We shall prove the equivalences of a non-degenerate circle-preserving map and a M(o)bius transformation in ^Rn, of a non-degenerate geodesic-preserving map and an isometry in Hn of a non-degenerate line-preserving map and an affine transformation in Rn. That a map is non-degenerate means that the image of the whole space under the map is not a circle, or geodesic or line respectively. These results hold without either injective or surjective, or even continuous assumptions, which are new and of a fundamental nature in geometry.

关键词： M(o)bius transformation, circle-preserving map, non-degenerate map.

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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 2012.3,china 3graduate school of the chinese academy of sciences,Beijing 1.0039,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.

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A Test on Different Types of the Time Curve of Hardness Ratio of Gamma-Ray Bursts based on the Curvature Effect

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chinese Journal of Astronomy and Astrophysics 2008年第4期8卷 451-464页

作者： Lan-Wei Jia National Astronomical Observatories/Yunnan Observatory,chinese academy of sciences,Kunming 650011,china graduate school of chinese academy of sciences,Beijing 100049,china National Astronomical Observatories/ Yunnan Observatory Chinese Academy of Sciences Kunming 650011 China graduate school of chinese academy of sciences Beijing 100049 China

We analyzed a sample of 66 gamma-ray bursts （GRBs） and statistically confirmed the prediction on the time curve of the hardness ratio of GRBs made by Qin et al. based on the curvature effect. In their analysis, GRB pulses are divided into three types according to the shape of their raw hardness ratio （RHR） time curves, defined as to include the background counts to the signal counts, so as to make use of counts within small time intervals. Of the three types, very hard sources exhibit a perfect pulse-like profile （type 1）, hard bursts possess a pulse-like profile with a dip in the decay phase （type 2）, and soft bursts show no pulse-like profile but have only a dipped profile （type 3）. In terms of the conventional hardness ratio, type 3 sources are indeed generally softer than those of type 1 and type 2, in agreement with the prediction. We found that the minimum value of RHR is sensitive in distinguishing the different types. We propose that GRB pulses can be classified according to the minimum value of RHR and that the different type sources may be connected with different strengths of the shock or/and the magnetic field.

关键词： gamma-rays burst -- gamma-rays observations - methods statistical

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Infrared Galaxies in the Nearby Universe

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chinese Journal of Astronomy and Astrophysics 2008年第6期8卷 643-652页

作者： Jian-Ling Wang 1 National Astronomical Observatories,chinese academy of sciences,Beijing 100012.china 2 graduate school of chinese academy of sciences,Beijing 100049,china National Astronomical Observatories Chinese Academy of Sciences Beijing 100012 China graduate school of chinese academy of sciences Beijing 100049 China

We used the Sloan Digital Sky Survey （SDSS） Data Release 5 （DR5） to study the morphological properties of 1137 nearby infrared （IR） galaxies, most of which are brighter than 15.9 mag in r-band. This sample was drawn from a cross-correlation of the Infra-Red Astronomical Satellite （IRAS） point source catalog redshift survey with DR5 at z ≤ 0.08. Based on this IR galaxy sample, we constructed five volume-limited sub-samples with IR luminosity ranging from 10^9.5 L⊙ to 10^12.⊙. By deriving the IR luminosity functions （LF） for different morphological types, we found that normal spiral galaxies are the dominant population below LIR ~ 8 ~ 10^10 L⊙; while the fraction of barred spiral galaxies increases with increasing IR luminosity and becomes dominant in spiral galaxies beyond LIR ≈ 5 × 10^10L⊙. As the IR luminosity decreases, the IR galaxies become more compact and have lower stellar masses. The analysis also shows that normal spiral galaxies give the dominant contribution to the total comoving IR energy density in the nearby universe, while, in contrast, the contribution from peculiar galaxies is only 39%.

关键词： galaxies evolution galaxies interactions galaxies starburst infrared galaxies galaxies luminosity function

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Effects of cosmological constant on motion of UHECR particles

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Science china(Physics,Mechanics & Astronomy) 2005年第1期48卷 44-56页

作者： CHEN Shaoxia & CHANG Zhe Institute of High Energy Physics, chinese academy of sciences, Beijing 1.0039, china graduate school of chinese academy of sciences, Beijing 1.0039, china Institute of High Energy Physics Chinese Academy of Sciences Beijing 100039 China graduate school Chinese Academy of Sciences Beijing 100039 China

Recent astronomical observations manifest that about two-thirds of the whole energy in the Universe is contributed by a small positive cosmological constant A (> 0). Then, an asymptotically de Sitter spacetime is premised naturally. However, physics in the de Sitter spacetime is very different from that in the Minkowski spacetime. As the first step, a covariant formalism of the kinematics in the de Sitter spacetime is presented here. By solving exactly the equations of motion for a field, we obtain the dispersion relation of a free particle. It is noticed that the dispersion relation is dependent on the degree of freedom of angular momentum of the particle. We show the threshold anomaly of the ultra high energy cosmic ray disappears naturally in the framework of the de Sitter kinematics.

关键词： ultrahigh energy cosmic ray, cosmological constant, curved spacetime.

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