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Identification and Characterization of glnA Promoter and its Corresponding Trans-regulatory Protein GlnR in the Rifamycin SV Producing Actinomycete,Amycolatopsis mediterranei U32

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Acta Biochimica et Biophysica Sinica 2006年第12期38卷 831-843页

作者： Hao YU~(1&) Wen-Tao peng~1 Yang LIU~1 Ting WU~2.Yu-Feng YAO~1 Ming-Xue CUI~1 Wei-Hong JIANG~1 Guo-Ping ZHAO~(1,3,4*) 1 laboratory of {2. Microbiology,institute of plant physiology and ecology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 2.0032.china 2.Department of Medicinal Chemistry,School of Pharmacy,Fudan University,shanghai 2.0032.china 3 State key laboratory of Genetic Engineering & Department of Microbiology,School of Life Science,Fudan University,shanghai 2.0433,china 4 laboratory of Disease and Health Genomics,chinese national Human Genome Center at shanghai,shanghai 2.12.3,china laboratory of molecular Microbiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China Department of Medicinal Chemistry School of Pharmacy Fudan University Shanghai 200032 China State key laboratory of Genetic Engineering & Department of Microbiology School of Life Science Fudan University Shanghai 200433 China laboratory of Disease and Health Genomics Chinese National Human Genome Center at Shanghai Shanghai 201203 China

The genetic requirements for the transcription of glnA,encoding the major glutaminesynthetase in a rifamycin SV-producing Amycolatopsis mediterranei strain,U32.were *** experiments showed that the promoter of U32.glnA （pglnA） was likely to have two transcriptioninitiation sites:P1and P2.located 157 and 45 nucleotides （nt） upstream of the translational start codon,*** mobility shift and DNase I footprinting analyses revealed a 30 bp cis-element locatedat 45 to 75 nt downstream of P1,or 38 to 68 nt upstream of *** sequence of the cis-element displayedhigh similarity to the corresponding regions of pglnA from Streptomyces coelicolor and *** as a reporter gene,the expression levels of U32.pglnA and its deletion derivatives under differentnitrogen-source conditions were analyzed by detecting the catechol dioxygenase activities in ***54,*** J508 and *** FS10 （glnR mutant）.These in vivo studies showed that theactivation of U32.pglnA in *** required GlnR,and its binding to the U32.pglnA was furtherconfirmed by the gel mobility shift *** and heterologous expression of the U32.glnR allowed usto detect the in vitro interaction between the U32.GlnR and the corresponding pglnA *** shown by in vivo glnR inactivation and complementation indicated that GlnR is essential for theactive transcription of glnA in U32.

关键词： glnA promoter cis-element glnR Amycolatopsis mediterranei

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Substitutions of the Conserved Gly47 Affect the CF1 Inhibitor and Proton Gate Functions of the Chloroplast ATP Synthase ε Subunit

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Acta Biochimica et Biophysica Sinica 2005年第7期37卷 453-462页

作者： Hui dong zhang-Lin NI Jia-Mian WEI {2. of plant physiology & ecology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai {12. institute of plant physiology and ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

The conserved residue Gly47 of the chloroplast ATP synthase ε subunit was substituted withLeu,Arg,Ala and Glu by site-directed *** process generated the mutants εG47L,εG47R,εG47A and εG47E,*** the ε variants showed lower inhibitory effects on the soluble CF1（-ε）Ca2.-ATPase compared with wild-type ε.In reduced conditions,εG47E and εG47R had a lower inhibitoryeffect on the oxidized CF1（-ε）Ca2.-ATPase compared with wild-type ε.In contrast,εG47L and εG47Aincreased the Ca2.-ATPase activity of soluble oxidized CF1（-ε）.The replacement of Gly47 significantlyimpaired the interaction between the subunit ε and γ in an in vitro binding *** study showed thatall ε variants were more effective in blocking proton leakage from the thylakoid *** enhancedATP synthesis of the chloroplast and restored ATP synthesis activity of the reconstituted membranes to alevel that was more efficient than that achieved by wild-type ε.These results indicate that the conservedGly47 residue of the ε subunit is very important for maintaining the structure and function of the ε subunitand may affect the interaction between the ε subunit,β subunit of CF1and subunit Ⅲ of CF_o,therebyregulating the ATP hydrolysis and synthesis,as well as the proton translocation role of the subunit Ⅲ of CF_o.

关键词： chloroplast ATP synthase ε subunit GST pull-down assay site-directed mutagenesis

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Thermostability of photosynthesis in two new chlorophyll b-less rice mutants

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Science china(Life sciences) 2005年第2期48卷 139-147页

作者： LIN Zhifang1,peng Changlian1,Xu Xinlan1,LIN Guizhu 1 & zhang Jingliu2 1.South china institute of Botany,chinese academy of sciences,Guangdong key laboratory of Digital Botanical Garden,Guangzhou 510650,china 2.national key laboratory of plant molecular genetics,institute of plant physiology & ecology,shanghai institutes of biological Sci- ences,chinese academy of sciences,shanghai 2.0032.china 1. South china institute of Botany Chinese Academy of Sciences Guangdong Key Laboratory of Digital Botanical Garden 510650 Guangzhou China 2. national key {2. of plant molecular genetics Institute of Plant Physiology & Ecology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences 200032 Shanghai China

Leaves of the two new chlorophyll b-less rice mutants VG2.-1, VG30-5 and the wild type rice cv. Zhonghua 11 were subjected to temperatures 2., 36, 40, 44 and 48℃ in the dark for 30 min or gradually elevated temperature from 30℃ to 80℃ at 0.5℃/min. The thermostability of photosynthetic apparatus was estimated by the changes in chlorophyll fluorescence parameters, photosynthetic rate and pigment content, chloroplast ultrastructure and tissue location of H2.2.accumulation. There were different patterns of Fo-temperature curves between the Chl b-less mutants and the wild type plant, and the temperature of Fo rising threshold was shifted 3℃ lower in the Chl b-less mutants (48℃) than in the wild type (51℃). At temperature up to about 45℃, chloroplasts were swollen and thylakoid grana became misty accompanied with the complete loss of photosynthetic oxygen evolution in the two Chl b-less mutants, but chloroplast ultrastruc- ture in the wild type showed no obvious alteration. After 55℃ exposure, the disordered thylakoid and significant H2.2.accumulation in leaves were found in the two Chl b-less mutants, whereas in the wild type plant, less H2.2.was accumulated and the swollen thylakoid still maintained a cer- tain extent of stacking. A large extent of the changes in qP, NPQ and Fv/Fm was consistent with the Pn decreasing rate in the Chl b-less mutants during high temperature treatment as compared with the wild type. The results indicated that the Chl b-less mutants showed a tendency for higher thermosensitivity, and loss of Chl b in LHC II could lead to less thermostability of PSII structure and function. Heat damage to photosynthetic apparatus might be partially attributed to the in- ternal oxidative stress produced at severely high temperature.

关键词： Oryza sativa L, thermostability, chlorophyll b-less mutant, chloroplast ultrastructure, chlorophyll fluorescence, H2.2.

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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 2.0032.china 2 Graduate School of the chinese academy of sciences,shanghai 2.0032.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 Lys2.,Lys101,Lys196,Lys2.7,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

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Cold stress-induced Transcriptome Changes in cassava(Manihot esculenta Crantz) by Long Oligonucleotide Microarray

Cold stress-induced Transcriptome Changes in cassava(Manihot...

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植物分子生物学与现代农业——2.10全国植物生物学研讨会

作者： dong An and peng zhang~* shanghai Center for Cassava Biotechnology,national laboratory of plant molecular genetics, institute of plant physiology & ecology,shanghai institutes for biological sciences,chinese academy of sciences,300 fenglin road,2.0032,shanghai,china.

Cassava is one of the most important tropical crops adapted to a wide range of environments,such as drought and infertile ***,it is categorized as extremely cold-sensitive *** our knowledge,little information is available about its cold signal reception and cold stress response although cold signaling pathway is well-investigated in model plant Arabidopsis *** development of

关键词： cassava long oligonucleotide microarray cold stress transcription factor families

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Functional study of CBF transcription factors from cassava (Manihot esculenta Crantz)

Functional study of CBF transcription factors from cassava (...

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上海市植物生理学会成立三十周年庆祝活动暨第三届上海市植物生理学青年学术研讨会

作者： dong An1 ,peng {2.2. SIBS-ETH shanghai Center for Cassava Biotechnology, national laboratory of plant molecular genetics, institute of plant physiology & {12., shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin road, shanghai 2.0032, china

DREB1/C-repeat binding factor (CBF) is a plant-specific family of transcription factors that play a crucial role in freezing tolerance[1,2.. In order to study the abiotic stress tolerance in cassava

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Functional Study of CBF Transcription Factors from Cassava （Manihot esculenta Crantz）

Functional Study of CBF Transcription Factors from Cassava ...

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中国植物生理学会第十次会员代表大会暨全国学术年会

作者： dong An, peng {2.* SIBS-ETH shanghai Center for Cassava Biotechnology, national laboratory of plant molecular genetics, institute of plant physiology & {12., shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin road, shanghai 2.0032, china

＜正＞DREB1/C-repeat binding factor （CBF） is a plant-specific family of transcription factors that play a crucial role

关键词： cassava CBF transcription factor abiotic stress tolerance gene function

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OXP1,a Vulnerable Protein to Oxidative Modification,Plays a Role in Starch Accumulation and Development in Arabidopsis

OXP1,a Vulnerable Protein to Oxidative Modification,Plays a ...

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植物分子生物学与现代农业——2.10全国植物生物学研讨会

作者： Xun-Liang Liu~1,Hai-dong Yu~1,Yuan Guan,Fang-Qing Guo~* The national {2. laboratory of plant molecular genetics,institute of plant physiology & ecology, shanghai institutes for biological sciences,{12.,300 fenglin road, shanghai2.0032.china

Carbonylation,an oxidative modification of proteins,is most commonly occurring during abiotic *** is thought to be an irreversible protein damage process and can be used as an overall protein oxidation *** modification usually leads to the loss of function of target proteins,which undergo proteolytic degradation or form

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Gene Expression Profile of Developing Cassava Storage Roots Revealed an Active Process of Glycolysis/Gluconeogenesis

Gene Expression Profile of Developing Cassava Storage Roots ...

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中国植物生理学会第十次会员代表大会暨全国学术年会

作者： Jun Yang, dong An, peng {2.* SIBS-ETH shanghai Center for Cassava Biotechnology, national laboratory of plant molecular genetics, institute of plant physiology & {12., shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin road, shanghai 2.0032, china

＜正＞Cassava storage root is a major sink for photosynthetic carbon assimilation. Its formation involves changes of transcriptional regulation and metabolic activities. For example, to accumulate starch in its parenc... 详细信息

关键词： cassava storage root development gene expression profile microarray

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Regulation of Starch Biosynthesis in Sweet potato [Ipomoea batatas （L.） Lam.] Towards Starch Quality Improvement

Regulation of Starch Biosynthesis in Sweet potato [Ipomoea b...

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中国植物生理学会第十次会员代表大会暨全国学术年会

作者： Jun Yang and peng zhang* SIBS-ETH {2. Center for Cassava Biotechnology, national laboratory of plant molecular genetics, institute of plant physiology & ecology,{12., chinese academy of sciences, 300 fenglin road, {2. 2.0032, china

＜正＞Granule-bound starch synthase I （GBSSI）, starch branching enzyme I （SBEI） and starch branching enzyme II （SBEII）, three key enzymes in starch biosynthesis, control starch composition in storage organs of p... 详细信息

关键词： sweet potato starch biosynthesis RNA interference genetic engineering

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