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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 200032,China institute of plant physiology & ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 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

Transcriptional, post-transcriptional and post-translational regulations of gene expression during leaf polarity formation

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Cell Research 2007年第6期17卷 512-519页

作者： Lin Xu Li YangHai Huang National 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 National Laboratory of plant Molecular Genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Present address: IBMP CNRS Universite Louis Pasteur 67084 Strasbourg France Present address: Department of Biology University of Pennsylvania Philadelphia PA 19104 USA

Leaf morphogenesis requires the establishment of adaxial-abaxial polarity after primordium initiation from the shootapical meristem (SAM). Several families of transcription factors are known to play critical roles in promoting adaxial orabaxial leaf fate. Recently, post-transcriptional gene silencing pathways have been shown to regulate the establishmentof leaf polarity, providing novel and exciting insights into leaf development. For example, microRNAs (miR165/166)and a trans-acting siRNA (TAS3-derived tasiR-ARF) have been shown to repress the expression of several key transcrip-tion factor genes. In addition, yet another level of regulation, post-translational regulation, has been revealed recently bystudies on the role of the 26S proteasome in leaf polarity. Although our understanding regarding the molecular mecha-nisms underlying establishment of adaxial-abaxial polarity has greatly improved, there is still much that remains *** review aims to discuss recent progress, as well as the remaining questions, regarding the molecular mechanismsunderlying leaf polarity formation.

关键词： Arabidopsis leaf development polarity formation

nifH Promoter Activity Is Regulated by DNA Supercoiling in Sinorhizobium meliloti

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 221-226页

作者： Yan-Jie LIU Biao HU Jia-Bi ZHU Shan-Jiong SHEN Guan-Qiao YU National Laboratory of plant Molecular Genetics,institute of plant physiology and ecology,shanghai institutes for Biological Sciences,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 Sciences Shanghai 200032 China

In prokaryotes,DNA supercoiling regulates the expression of many genes;for example,theexpression of Klebsiella pneumoniae nifLA operon depends on DNA negative supercoiling in anaerobicallygrown cells,which indicates that DNA supercoiling might play a role in gene regulation of the *** the expression of the nifH promoter in Sinorhizobium meliloti is not repressed by oxygen,itis proposed that the status of DNA supercoiling may not affect the expression of the nifH *** this hypothesis by analyzing nifH promoter activity in wild-type and gyr Escherichia coli in thepresence and absence of DNA gyrase *** results show that gene expression driven by the *** nifH promoter requires the presence of active DNA *** DNA gyrase increases thenumber of negative superhelical turns in DNA in the presence of ATP,our data indicate that negativesupercoiling is also important for nifH promoter *** study also shows that the DNA supercoiling-dependent *** nifH promoter activity is related to the trans-acting factors NtrC and NifA that *** supercoiling appeared to have a stronger effect on NtrC-activated nifH promoter activity than onNifA-activated promoter ***,these results from the *** nifH promoter model systemseem to indicate that,in addition to regulating gene expression during anaerobic signaling,DNA supercoilingmay also provide a favorable topology for trans-acting factor binding and promoter activation regardless ofoxygen status.

关键词： nifH DNA supercoiling gyrase Sinorhizobium meliloti gene expression

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, 200032 shanghai, China 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 el Max-Planck-institute of Molecular plant physiology (MPI-MP) on “plant Molecular physiology and Signal Transduction” 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.

Interaction of rice bZIP protein REB with the 5'-upstream region of both rice sbe1 gene and waxy gene

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Chinese Science Bulletin 2002年第4期47卷 310-314页

作者： CAI Yi, XIE Donglu, WANG Zongyang & HONG MengminState Key Laboratory of plant Molecular Genetics, shanghai institute of plant physiology and ecology, Chinese Academy of Sciences, shanghai 200032, China State Key Laboratory of plant Molecular Genetics Shanghai Institute of Plant Physiology and Ecology Chinese Academy of Sciences Shanghai China

Rice starch branching enzyme 1 (SBE1) and granule-bound starch synthase (GBSS) catalyzed the bio-synthesis of amylopectin and amylose in rice developing en-dosperm respectively, and the genes encoding these two en-zymes, sbe1 and waxy, are mainly expressed in the en-dosperm. Within the 5 ’-upstream region of rice sbel gene, we identified a 53 bp fragment C53 which could interact with the nuclear proteins extracted from rice endosperm. We also found that fragment Ha-2 in the 5 ’-upstream region of rice waxy gene could compete with C53 for its interaction. Fur-ther experiments demonstrated that rice bZIP protein REB could interact with two ACGT elements (G-box and Hex) in the fragment C53 as well as three ACGT elements (WG1, WG2, and WG3) in the fragment Ha-2. The WG1 element could compete well with Hex element for its interaction with REB. These results suggested that the expression of sbel and waxy gene in rice developing endosperm may be coordinately regulated by REB-like bZIP transcriptional factors.

关键词： rice waxy gene sbel gene bZIP protein.

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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, 200032 shanghai, China National Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences (SIBS) Chinese Academy of Sciences Partner Group of Max-Planck-institute of Molecular plant physiology (MPI-MP) on “plant Molecular physiology and Signal Transduction” Shanghai 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.

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 200032,China institute of plant physiology and ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Graduate School of the Chinese Academy of SciencesShanghai 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

Identification of ABA-responsive genes in rice shoots via cDNA macroar- ray

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Cell Research 2003年第1期13卷 59-68页

作者： FANG LIN, SHOU LING XU, WEI MIN NI, ZHAO QING CHU, ZHI HONG XU, HONG WEI XUENational Laboratory of plant Molecular Genetics, institute of plant physiology and ecology, shanghai institutes for Biological Science (SIBS), Chinese Academy of Sciences and Partner Group of Max-Planck-institute of Molecular plant physiology (MPI-MP) on "plant Molecular physiology and Signal Transduc-tion", 300 Fenglin Road, 200032 shanghai, China National Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Science (SIBS) Chinese Academy of Sciences Partner Group of Max-Planck-institute of Molecular plant physiology (MPI-MP) on plant Molecular physiology and Signal Transduction Shanghai China

Phytohormone abscisic acid (ABA) was critical for many plant growth and developmental processes including seed maturation, germination and response to environmental factors. With the purpose to detect the possible ABA related signal transduction pathways, we tried to isolate ABA-regulated genes through cDNA macroarray technology using ABA-treated rice seedling as materials (under treatment for 2, 4, 8 and 12 h). Of 6144 cDNA clones tested, 37 differential clones showing induction or suppression for at least one time, were isolated. Of them 30 and 7 were up- or down-regulated respectively. Sequence analyses revealed that the putative encoded proteins were involved in different possible processes, including transcription, metabolism and resistance, photosynthesis, signal transduction, and seed maturation. 6 cDNA clones were found to encode proteins with unknown functions. Regulation by ABA of 7 selected clones relating to signal transduction or metabolism was confirmed by reverse transcription PCR. In add

关键词： Oryza saliva abscisic acid (ABA) cDNA macroarray responsive genes signal transduc- tion

Are small RNAs a big help to plants?

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Science China(Life Sciences) 2009年第3期52卷 212-223页

作者： MAO YingBo,XUE XueYi & CHEN XiaoYa National Key Laboratory of plant Molecular Genetics,institute of plant physiology and ecology,shanghai institutes for Biological Sciences,Chinese Academy of Sciences,shanghai 200032,China National Key Laboratory of plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

The discovery of RNA interference(RNAi) has augmented our knowledge of gene regulation and presents a fascinating technology that has a great potential for application in genetic analysis,disease therapy,plant protection,and many other *** this review,we will focus on the biological functions of RNAi and its application in agriculture with a brief introduction to the history of its discovery and molecular mechanism.

关键词： RNA interference gene regulation plant protection

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