Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Proteomic Analysis of Cytokinin Induced Proteins in Arabidopsis

단백체를 이용한 애기장대 Cytokinin 유도 단백질의 분석

  • Liang Ying-Shi (Division of Applied Life Science, Gyeongsang National University, PMBBRC, Gyeongsang National University) ;
  • Cha Joon-Yung (Division of Applied Life Science, Gyeongsang National University, PMBBRC, Gyeongsang National University, EBNCRC, Gyeongsang National University) ;
  • Ermawati Netty (Division of Applied Life Science, Gyeongsang National University, PMBBRC, Gyeongsang National University) ;
  • Jung Min-Hee (Division of Applied Life Science, Gyeongsang National University, PMBBRC, Gyeongsang National University) ;
  • Bae Dong-Won (Central Laboratory, Gyeongsang National University) ;
  • Lee Chang-Won (Department of Microbiology, Gyeongsang National University) ;
  • Son Dae-Young (Division of Applied Life Science, Gyeongsang National University, PMBBRC, Gyeongsang National University, EBNCRC, Gyeongsang National University)
  • 양영실 (경상대학교 대학원 응용생명과부, 식물분자생물학 및 유전자조작연구소) ;
  • 차준영 (경상대학교 대학원 응용생명과부, 식물분자생물학 및 유전자조작연구소, 환경생명과학 국가핵심 연구센터) ;
  • 네티 엘마와티 (경상대학교 대학원 응용생명과부, 식물분자생물학 및 유전자조작연구소) ;
  • 정민희 (경상대학교 대학원 응용생명과부, 식물분자생물학 및 유전자조작연구소) ;
  • 배동원 (경상대학교 공동실험실습관) ;
  • 이창원 (경상대학교 미생물학과) ;
  • 손대영 (경상대학교 대학원 응용생명과부, 식물분자생물학 및 유전자조작연구소, 환경생명과학 국가핵심 연구센터)
  • Published : 2005.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. To better understand the molecular mechanisms of cytokinin action, we identified cytokinin related proteins by a proteomic approach. Proteins extracted from control and trans-zeatin treated Arabidopsis seedlings were separated and analyzed by two dimensional gel analysis. Differentially expressed protein spots were identified with peptide mass fingerprinting based on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and database searching, We obtained ten up-regulated and one down-regulated proteins upon t-zeatin treatment. The expression of the following proteins was induced; pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase and IAA amino acid hydrolase homolog.

Cytokinin은 식물의 성장과 발달에 중요한 역할을 하는 필수 호르몬이다. Cytokinin의 작용 기작을 이해하기 위하여 단백체를 이용하여 cytokinin 관련 단백질들을 동정하였다. 대조구와 t-zeatin을 처리한 애기장대로부터 추출한 단백질을 이차원 전기영동하여 분석하였다. 발현양에 차이가 있는 단백질 spot들을 MALDI-TOF 단백질 질량분석기와 database 검색을 통하여 동정하였다. 그 결과 t-zeatin 처리에 의하여 발현이 증가하는 10개의 단백질과 감소하는 한 개의 단백질을 분리할 수 있었다. Cytokinin에 의하여 발현이 증가하는 단백질은 pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase, 그리고 IAA amino acid hydrolase homolog들로 밝혀졌다.

Keywords

References

  1. Bianchi MW, Roux C, Vartanian N (2002) Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase. Physiol Plant 116: 96-105 https://doi.org/10.1034/j.1399-3054.2002.1160112.x
  2. Binns AN (1994) Cytokinin accumulation and action: biochemical, genetic and molecular approaches. Annnu Rev Plant Physiol Plant Mol Biol 45: 197-209 https://doi.org/10.1146/annurev.pp.45.060194.001213
  3. Blum H, Beier H, Gross HJ (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8: 93-99 https://doi.org/10.1002/elps.1150080203
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  5. Crowell DN (1994) Cytokinin regulation of a soybean pollen allergen gene. Plant Mol Biol 25: 829-835 https://doi.org/10.1007/BF00028877
  6. Durmus N, Kadioglu A (2005) Reduction of paraquat toxicity in maize leaves by benzyladenine. Acta Biol Hung 56: 97-107 https://doi.org/10.1556/ABiol.56.2005.1-2.10
  7. Flores S, Tobin EM (1989) Cytokinin modulation of LHCP mRNA levels: the involvement of post-transcriptional regulation. Plant Mol Biol 11: 409-415 https://doi.org/10.1007/BF00039021
  8. Howell SH, Lall S, Che P (2003) Cytokinins and shoot development. Trends Plant Sci 8: 453-459 https://doi.org/10.1016/S1360-1385(03)00191-2
  9. Jensen ON, Wilm M, Shevchenko A, Mann M. (1999) Sample preparation methods for mass spectrometric peptide mapping directly from 2-DE gels. Methods Mol Biol 112: 513-530
  10. Kakimoto T (1996) CKI1, a histidine kinase homolog implicated in cytokinin signal transduction. Science 274: 982-985 https://doi.org/10.1126/science.274.5289.982
  11. Kilili KG, Atanassova N, Vardanyan A, Clatot N, Al-Sabarna K, Kanellopoulos PN, Makris AM, Kampranis SC (2004) Differential roles of tau class glutathione S-transferases in oxidative stress. J Biol Chem 279: 24540-24551 https://doi.org/10.1074/jbc.M309882200
  12. Kim ST, Cho KS, Jang YS, Kang KY (2001) Two-dimensional electrophoretic analysis of rice proteins by polyethylene glycol fractionation for protein arrays. Electrophoresis 22: 2103-2109 https://doi.org/10.1002/1522-2683(200106)22:10<2103::AID-ELPS2103>3.0.CO;2-W
  13. Landry F, Lombardo CR, Smith JW (2000) A method for application of samples to matrix-assisted laser desorption ionization time-of-flight targets that enhances peptide detection. Anal Biochem 279: 1-8 https://doi.org/10.1006/abio.1999.4468
  14. Lebs S, Lebs W, Klyachko NL, Romanko EG, Kulaeva ON, Wollgiehn R, Parthier B (1984) Gene expression in cytokinin- and light-mediated plastogenesis of cucurbita cotylendons: ribulose-l ,5-bisphosphate carboxylase/oxygenase. Planta 162: 289-298 https://doi.org/10.1007/BF00396739
  15. Longo GP, Bracale M, Rossi G, Longo CP (1990) Benzyladenine induces the appearance of LHCP-mRNA and of the relevant protein in dark-grown excised watermelon cotyledons. Plant Mol Biol 14: 569-573 https://doi.org/10.1007/BF00027502
  16. Lu JL, Ertl JR, Chen CM (1990) Cytokinin enhancement of the light incubation of nitrate reductase transcript levels in etiolated barley leaves. Plant Mol Biol 14: 585-594 https://doi.org/10.1007/BF00027504
  17. Menges M, Hennig L, Gruissem W, Murray JA (2002) Cell cycle regulated gene expression in Arabidopsis. J Biol Chem 277: 41987-42002 https://doi.org/10.1074/jbc.M207570200
  18. Mok DW, Mok MC (2001) Cytokinin metabolism and action. Annu Rev Plant Physiol Plant Mol Biol 52: 89-118 https://doi.org/10.1146/annurev.arplant.52.1.89
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15: 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  20. Muskett P, Parker J (2003) Role of SGT1 in the regulation of plant R gene signalling. Microbes Infect 5: 969-976 https://doi.org/10.1016/S1286-4579(03)00183-7
  21. O’Farrell, PH (1975) High resolution two-dimensional electrophoresis of protein. J Biol Chem 250: 4007-4021
  22. Rashotte AM, Carson SD, To JP, Kieber JJ (2003) Expression profiling of cytokinin action in Arabidopsis. Plant Physiol 132: 1998-2011 https://doi.org/10.1104/pp.103.021436
  23. Rupp HM, Frank M, Werner T, Strnad M, Schmulling T (1999) Increased steady state mRNA levels of the STM and KNATI homeobox genes in cytokinin overproducing Arabidopsis thaliana indicate a role for cytokinins in the shoot apical meristem. Plant J 18: 557-563 https://doi.org/10.1046/j.1365-313X.1999.00472.x
  24. Suzuki I, Cretin C, Omata T, Sugiyama T (1994) Transcriptional and posttranscriptional regulation of nitrogen-responding expression of phosphoenol-pyurvate carboxylase gene in maize. Plant Physiol 105: 1223-1229 https://doi.org/10.1104/pp.105.4.1223