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

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.

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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, 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.

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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, 2.0032 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

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MORN motifs in plant PIPKs are involved in the regulation of subcellular localization and phospholipid binding

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Cell Research 2006年第5期16卷 466-478页

作者： Hui Ma~(1,*) Ying Lou~1 Wen Hui Lin~1 Hong Wei Xue~1 ~1National Key Laboratory of plant molecular Genetics,institute of plant physiology and Ecology,shanghai institutes for Biological Science(SIBS),Chinese Academy of Sciences and Parmer 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 Key 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 transduction” 300 Fenglin Road 200032 Shanghai China 1National Key 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 transduction” 300 Fenglin Road 200032 Shanghai China 1National Key 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 transduction” 300 Fenglin Road 200032 Shanghai China 1National Key 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 transduction” 300 Fenglin Road 200032 Shanghai China

Multiple repeats of membrane occupation and recognition nexus （MORN） motifs were detected in plant phosphati-dylinositl monophosphate kinase （PIPK）,a key enzyme in PI-signaling *** analysis indicates that allthe MORN motifs （with varied numbers at ranges of 7-9）,which shared high homologies to those of animal ones,werelocated at N-terminus and sequentially arranged,except those of OsPIPK1 and AtPIPK7,in which the last MORN motifwas separated others by an～100 amino-acid"island"region,revealing the presence of two kinds of MORN arrangementsin plant *** employing a yeast-based SMET （sequence of membrane-targeting） system,the MORN motifswere shown being able to target the fusion proteins to cell plasma membrane,which were further confirmed by expressionof fused MORN-GFP *** detailed analysis via deletion studies indicated the MORN motifs in OsPIPK1,together with the 104 amino-acid"island"region are involved in the regulation of differential subcellular localization,*** membrane or nucleus,of the fused *** Western blot analysis of the recombinant MORN polypep-tide,expressed in Escherichia coli,showed that MORN motifs could strongly bind to PA and relatively slightly to PI4Pand PI（4,5）*** results provide informative hints on mechanisms of subcellular localization,as well as regulationof substrate binding,of plant PIPKs.

关键词： membrane targeting PIPK MORN motif subcellular localization

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水稻eIF3大亚基(eIF3a)编码基因的克隆及其表达模式分析

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实验生物学报 2003年第1期36卷 54-60页

作者：李竑沈思师许智宏薛红卫上海师范大学中国科学院上海生命科学研究院植物生理研究所植物分子遗传国家重点实验室

真核生物翻译起始因子(eIFs)在蛋白合成中起关键作用,在已经鉴定的13个因子中eIF3(由8个或更多的亚基组成)是分子量最大的一个并在翻译起始过程中起着核心作用。eIF3a是eIF3中最大的亚基并介导了eIF3的大多数的功能的实现。利用荧光-差... 详细信息

真核生物翻译起始因子(eIFs)在蛋白合成中起关键作用,在已经鉴定的13个因子中eIF3(由8个或更多的亚基组成)是分子量最大的一个并在翻译起始过程中起着核心作用。eIF3a是eIF3中最大的亚基并介导了eIF3的大多数的功能的实现。利用荧光-差异显示PCR法对生长素处理后的水稻材料进行分析发现eIF3a的表达可被生长素诱导,进一步通过筛选cDNA文库分离出水稻编码eIF3a的全长cDNA(命名为OseIF3a1),OseIF3a1含3459碱基(含5’和3’非翻译区)并编码一个986氨基酸的蛋白,与基因组序列比较表明OseIF3a1基因存在有12个内含子。OSeIF3a1与玉米和烟草的同源蛋白一致性分别为82.4%和70.1%并与其他真核生物eIF3a蛋白具较高一致性。以组织材料进行的RT-PCR结果表明OseIF3a1在根、幼苗、幼穗、茎和叶中表达,启动子-报告基因的转基因结果进一步表明OseIF3a1在根尖及幼嫩叶片表达较高,进一步的RTPCR分析确证了生长素对OseIF3a1的诱导,表明生长素在调控植物生长时可能涉及了翻译水平上的调节。

关键词： eIF3大亚基编码基因水稻克隆表达模式真核翻译起始因子荧光差异显示PCR 生长素

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