Molecular analysis of a MAP kinase cascade involved in environment stress response in Arabidopsis thaliana 環境ストレス応答におけるシロイヌナズナMAPキナーゼカスケードに関する分子生物学的解析
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著者
書誌事項
- タイトル
-
Molecular analysis of a MAP kinase cascade involved in environment stress response in Arabidopsis thaliana
- タイトル別名
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環境ストレス応答におけるシロイヌナズナMAPキナーゼカスケードに関する分子生物学的解析
- 著者名
-
市村, 和也
- 著者別名
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イチムラ, カズヤ
- 学位授与大学
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筑波大学
- 取得学位
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博士 (理学)
- 学位授与番号
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甲第2016号
- 学位授与年月日
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1999-03-25
注記・抄録
博士論文
1998
【要旨】
目次
- Contents / p1 (0003.jp2)
- Abbreviations / p4 (0006.jp2)
- Abstract / p1 (0008.jp2)
- Introduction / p8 (0015.jp2)
- Materials and Methods / p18 (0025.jp2)
- Plant materials / p19 (0026.jp2)
- Strains,media and yeast transformation / p19 (0026.jp2)
- Molecular biological techniques / p19 (0026.jp2)
- Two-hybrid screening / p20 (0027.jp2)
- Molecular cloning of a full-length cDNA for ATMKK2 / p21 (0028.jp2)
- Preparation of cDNA fragments for ATMKK3,4 and 5 for cDNA library screening / p21 (0028.jp2)
- Cloning and sequencing of cDNAs for ATMKK3,4 and 5 / p22 (0029.jp2)
- Southern and Northern blot analysis / p23 (0030.jp2)
- Expression of GST-fusion proteins in E. coli and phosphorylation assay of GST-fusion proteins / p23 (0030.jp2)
- Mapping of the ATMKK2,3,4 and 5 genes / p24 (0031.jp2)
- Cloning of ANP4 / p24 (0031.jp2)
- Two-hybrid and β-galactosidase assays / p25 (0032.jp2)
- Complementation of the growth defect of the pbs2Δ mutant by coexpression of Arabidopsis MAPKKs and MAPKKKs / p25 (0032.jp2)
- Complementation of the growth defects of the mpk1Δ and the bck1Δ mutants by coexpression of Arabidopsis MAPKs and MAPKKs / p26 (0033.jp2)
- Stress and phytohormone treatment / p26 (0033.jp2)
- Preparation of protein extracts / p27 (0034.jp2)
- Protein kinase assay / p27 (0034.jp2)
- Immunoprecipitation / p28 (0035.jp2)
- Antibody production and immunoblot analysis / p28 (0035.jp2)
- Results / p30 (0037.jp2)
- Part1:Isolation of ATMEKK1-interacting proteins using a yeast two-hybrid system / p31 (0038.jp2)
- Isolation of ATMEKK1-interacting proteins using a yeast two-hybrid system / p31 (0038.jp2)
- Nucleotide and amino acid sequence,and expression of ATMKK2(a MAPKK) / p33 (0040.jp2)
- Part2:Isolation and characterization of Arabidopsis MAPKK homologues / p34 (0041.jp2)
- Structural analysis of the putative ATMKK2,3,4 and 5 proteins / p35 (0042.jp2)
- Detection of protein kinase activity of the recombinant ATMKK3 and 4 proteins / p37 (0044.jp2)
- Northern and Southern blot analysis of the ATMKK3,4,and 5 genes / p38 (0045.jp2)
- Map positions of the ATMKK2,3,4 and 5 genes on Arabidopsis chromosomes / p39 (0046.jp2)
- Part3:Identification of a possible MAPK cascade based on yeast two-hybrid analysis and complementation of yeast mutants / p40 (0047.jp2)
- Analysis of complex formation between Arabidopsis MAPKKKs and MAPKKs using the yeast two-hybrid system / p40 (0047.jp2)
- Analysis of complex formation between Arabidopsis MAPKKs and MAPKs using the yeast two-hybrid system / p41 (0048.jp2)
- Analysis of complex formation between Arabidopsis MAPKs and MAPKKKs using the yeast two-hybrid system / p42 (0049.jp2)
- Deletion mappings of ATMPK4(a MAPK) and MEK1/ATMKK2(MAPKKs)interacting domains of ATMEKK1(a MAPKKK) / p43 (0050.jp2)
- Expression of ATMEKK1(a MAPKKK)increases activity of two closely related MAPKKs,MEK1 and ATMKK2,to complement the pbs2Δ defect / p44 (0051.jp2)
- Substitution of either threonine residue 220 or 226 with alanine completely abolishes the ability of A TMKK2 to suppress the pbs2 mutant / p45 (0052.jp2)
- Coexpression of MEK1(a MAPKK)and ATMPK4(a MAPK)complements the bck1Δ and mpk1Δ defects / p46 (0053.jp2)
- Part4: Effect of environmental stresses on the activation of Arabidopsis MAP kinase,ATMPK4 / p49 (0056.jp2)
- The ATMPK4CT antibody specifically recognize the ATMPK4 protein / p49 (0056.jp2)
- ATMPK4 is transiently activated by cold,dehumidification,touch and wounding stress treatments / p50 (0057.jp2)
- ATMPK4 is not activated by heat shock and ABA,partially activated by MeJA / p52 (0059.jp2)
- Low temperature but not osmotic shock,heat shock or ABA,induced activation of ATMPK4 in T87 cultured cell / p52 (0059.jp2)
- Expression of the ATMPK4 gene did not correlate with activity of ATMPK4 protein under stress conditions / p54 (0061.jp2)
- Protein level of ATMPK4 does not co-relate with its activity under stress conditions / p55 (0062.jp2)
- Discussion / p56 (0063.jp2)
- Isolation of ATMEKK1 binding proteins using yeast two-hybrid system / p57 (0064.jp2)
- Physical interactions among ATMEKK1(MAPKKK),MEK1/ATMKK2(MAPKKs) and ATMPK4(MAPK) / p58 (0065.jp2)
- Functional interactions among ATMEKK1(MAPKKK),MEK1/ATMKK2(MAPKKs) and ATMPK4(MAPK) / p61 (0068.jp2)
- ATMPK4,an Arabidopsis MAPK homologue,is activated by cold,dehumidification,touch and wounding stresses / p62 (0069.jp2)
- Possible roles of ATMPK4 kinase under cold and drought stress conditions / p63 (0070.jp2)
- A possible role of ATMPK4 under mechanical stress con ditions / p64 (0071.jp2)
- A possible role of ATMPK4 under wounding stress conditions / p65 (0072.jp2)
- Relationships between ATMPK4 and stress-activated MAPKs in plants / p66 (0073.jp2)
- A possible function of stress-activated ATMPK4-MAPK cascade in stress response / p68 (0075.jp2)
- Conclusions and perspectives / p71 (0078.jp2)
- Figures and Tables / p74 (0081.jp2)
- References / p123 (0130.jp2)
- Acknowledgements / p147 (0154.jp2)