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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 institute of plant physiology & ecology.shanghai institutes for biological sciences.chinese academy of sciences.shanghai 2.0032.china 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

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 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

The tricks plants use to reach appropriate light

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Science china(Life sciences. 2010年第8期53卷 916-926页

作者： CHAO DaiYin1 & LIN HongXuan2 1Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907, USA 2National Key Laboratory of plant Molecular Genetics, shanghai institute of plant physiology and ecology. shanghai institutes for biological sciences. chinese academy of sciences. shanghai 2.0032. china 1. Department of Horticulture and Landscape Architecture Purdue University West Lafayette Indiana 47907 USA2. National Key Laboratory of Plant Molecular Genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

The perception of ambient light signals that produce a relevant response to ensure exposure to appropriate levels of light energy is vital for plants. In response to this, intricate molecular mechanisms to mediate light signaling have evolved in plants. Among the responses induced by light, seedling extension is a determining event for plant survival in darkness, especially in the initial stage of plant growth. Here we review previous studies and recent progress towards an understanding of light signaling that regulates seedling elongation. We focus on the three regions of the sunlight spectrum that primarily control seedling elongation, namely red/far-red light, blue/UV-A light and UV-B light, and summarize the four signaling pathways that correspond to the three effective spectra.

关键词： photomorphogenesis light signaling photoreceptors cell elongation cell division

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HAL1 mediate salt adaptation in Arabidopsis thaliana

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Cell Research 2001年第2期11卷 142-148页

作者： YANG SU XIN, YAN XIU ZHAO, QUAN ZHANG, YU KE HE, HUI ZHANG, DA LUO institute of plant physiology, shanghai institutes for biological sciences. chinese academy of Science, 300 Fenglin Road, shanghai 2.0032. china Key Laboratory of plant Stress Research, Sh 1institute of plant physiology Shanghai Institutes for Biological Sciences Chinese Academyof Science 300 Fenglin Road Shanghai 200032 China 2Key Laboratory of plant Stress Research Shandong Normal University Jinan 250014 China 2Key Laboratory of plant Stress Research Shandong Normal University Jinan 250014 China 1institute of plant physiology Shanghai Institutes for Biological Sciences Chinese Academyof Science 300 Fenglin Road Shanghai 200032 China 2Key Laboratory of plant Stress Research Shandong Normal University Jinan 250014 China 1institute of plant physiology Shanghai Institutes for Biological Sciences Chinese Academyof Science 300 Fenglin Road Shanghai 200032 China

The yeast HAL1 gene was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens-mediated transformation with vacuum infiltration under the control of CaMV 35S promoter. Thirty-three individual kanamycin resistant plants were obtained from 75,000 seeds. Southern blotting analysis indicated that HAL1 gene had been integrated into all of the transgenic plants’ genomes. The copy number of HAL1 gene in transgenic plants was mostly 1 to 3 by Southern analysis. Phenotypes of transgenic plants have no differences with wild type plants. several samples of transformants were self-pollinated, and progenies from transformed and non-transformed plants (controls) were evaluated for salt tolerance and gene expression. Measurement of concentrations of intracellular K+ and Na+ showed that transgenic lines were able to retain less Na+ than that of the control under salt stress. Results from different tests indicated the expression of HAL1 gene promotes a higher level of salt tolerance in vivo in the transgenic Arabidopsis plants.

关键词： Salt-tolerant Arabidopsis HAL1 gene engineering.

In Vitro Reassembly of Tobacco Ribulose-1,5-bisphosphate Carboxylase/Oxygenase from Fully Denatured Subunits

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Acta Biochimica et Biophysica Sinica 2006年第10期38卷 737-745页

作者： Zhen-Hua YONG Gen-Yun CHEN* Jiao-Nai SHI Da-Quan XU institute of plant physiology and ecology.shanghai institutes for biological sciences.chinese academy of sciences.and Graduate School of the chinese academy of sciences.shanghai 2.0032.china *** institute of plant physiology and ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Graduate School of the Chinese Academy of Sciences Shanghai 200032 China

It has been generally proved impossible to reassemble ribulose-1,5-bisphosphate carboxylase/oxygenase （Rubisco） from fully denatured subunits in vitro in higher plant,because large subunit of fullydenatured Rubisco is liable to precipitate when the denaturant is removed by common methods of directdilution and one-step *** our experiment,the problem of precipitation was resolved by an improvedgradual dialysis method,which gradually decreased the concentration of ***,fully denaturedRubisco subunits still could not be reassembled into holoenzyme using gradual dialysis unless chaperonin 60was *** restored activity of reassembled Rubisco was approximately 8% of natural *** of reassembled Rubisco increased greatly when heat shock protein 70 was present in the *** and Mg2+ were unnecessary for in vitro reassembly of Rubisco,and Mg2+ inhibited the *** reassembly was weakened when ATP,Mg2+ and K+ existed together in the reassembly process.

关键词： Rubisco reassembly gradual dialysis chaperonin 60 Hsp70

DNA chip-based expression profile analysis indicates involvement of the phosphatidylinositol signaling pathway in multiple plant responses to hormone and abiotic treatments

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Cell Research 2004年第1期14卷 34-45页

作者： Wen Hui LIN, Rui YE, Hui MA, Zhi Hong XU, Hong Wei XUE National Laboratory of plant Molecular Genetics, institute of plant physiology and ecology. shanghai institutes for biological sciences.(SIBS), chinese academy of sciences.and Partner Group of Max-Planck-institute of Molecular plant physiology (MPI-MP) on "plant Molecular physiology and Signal Transduction", 300 Fenglin Road, 2.0032.shanghai, china Xue HW Chinese Acad Sci Shanghai Inst Biol Sci Inst Plant Physiol & Ecol Natl Lab Plant Mol Genet 300 Fenglin Rd Shanghai 200032 Peoples R China

The phosphatidylinositol (PI) metabolic pathway is considered critical in plant responses to many environmental factors, and previous studies have indicated the involvement of multiple PI-related gene families during cellular responses. Through a detailed analysis of the Arabidopsis thaliana genome, 82 polypeptides were identified as being involved in PI signaling. These could be grouped into different families including PI synthases (PIS), PI-phosphate kinases (PIPK), phospholipases (PL), inositol polyphosphate phosphatases (IPPase), inositol polyphosphate kinases (IPK), PI transfer proteins and putative inositol polyphosphate receptors. The presence of more than 10 isoforms of PIPK, PLC, PLD and IPPase suggested that these genes might be differentially expressed during plant cellular responses or growth and development. Accordingly, DNA chip technology was employed to study the expression patterns of various isoforms. In total, 79 mRNA clones were amplified and used for DNA chip generation. Expression

关键词： phosphatidylinositol signal transduction DNA chip expression profile.

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 2.0032. china. ~2Biotechnology institute, Zhejiang University, Hangzhou 310029, china. 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. 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. 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.

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.

Isolation and functional characterization of the C-terminus of rice phos-phatidylinositol 4-kinase in vitro

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Cell Research 2003年第2期13卷 131-139页

作者： XIANG FENG KONG, ZHI HONG XU, HONG WEI XUE, National Laboratory of plant Molecular Genetics, institute of plant physiology and ecology. shanghai institutes for biological Science (SIBS), The chinese academy of sciences.Partner Group of Max-Planck-institute of Molecular plant physiology (MPI-MP) on "plant Molecular physiology and Signal Transduction", 300 Fenglin Road, 2.0032.shanghai, china 1National Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Science (SIBS)The Chinese Academy of Sciences Partner Group el Max-Planck-institute of Molecular plant physiology (MPI-MP) on “plant Molecular physiology and Signal Transduction” 300 Fenglin Road 200032 Shanghai China 1National Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Science (SIBS)The Chinese Academy of Sciences Partner Group el Max-Planck-institute of Molecular plant physiology (MPI-MP) on “plant Molecular physiology and Signal Transduction” 300 Fenglin Road 200032 Shanghai China 1National Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Science (SIBS)The Chinese Academy of Sciences Partner Group el Max-Planck-institute of Molecular plant physiology (MPI-MP) on “plant Molecular physiology and Signal Transduction” 300 Fenglin Road 200032 Shanghai China

A partial rice (Oryza sativa L.) cDNA clone. OsPMK1c, was isolated through screening of a cDNA library constructed from tillering materials. OsPI4Klc encoded a peptide of 608 amino acids with a calculated molecular mass of 68.4 kDa. The OsPI4Klc peptide shared high homology and possessed the highly conserved domains present in most isolated cloned PI4-kinases, i.e. a lipid kinase unique (LKU) domain and a catalytic (CAT) domain. A region with similarity to pleckstrin homology (PH) domain was present in OsPI4K1c as well. Further comparison with genomic sequences in databases revealed that OsPI4K1c is located at the 3'-end of a putative rice PI 4-kinase coding gene OsPI4K1, and its coding region corresponded to the C-terminal half of OsPI4Kl protein. Twelve exons (49-562 bp in size) and 11 introns (77-974 bp in size) were identified in OsPI4K1c. The recombinant protein expressed in Escherichia coli phosphorylates phosphatidylinositol at the D4 position of the inositol ring. OsPI4K1 transcript levels were

关键词： Oryza sativa phosphatidylinositol 4-kinase.

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 2.0032. china, 2SHARF Laboratory, shanghai institute of plant physiology and ecology. shanghai institutes for biological sciences. chinese academy of sciences. 300 Fenglin Road, shanghai 2.0032. 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.