Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Identification of Brassinosteroid-Related Protein, BAK1 from Nutrition Deficient Tomato Cultivated by Soilless Cultivation System

수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정

  • Shin, Pyung-Gyun (Plant Nutrition Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Chang, An-Cheol (Plant Nutrition Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Hong, Sung-Chang (Plant Nutrition Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Lee, Ki-Sang (Plant Nutrition Division, National Institute of Agricultural Science and Technology, RDA)
  • 신평균 (농업과학기술원 식물영양과) ;
  • 장안철 (농업과학기술원 식물영양과) ;
  • 홍성창 (농업과학기술원 식물영양과) ;
  • 이기상 (농업과학기술원 식물영양과)
  • Published : 2007.12.30
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Abstract

Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

BAK1(Brassinolide insensitive associated receptor kinase 1)는 브라시노스테로이드 생합성 대사관련 신호전달 매체이다. BR 생합성 및 신호전달 돌연변이체는 매우 특징적인 난쟁이 표현형을 보인다. 과채류전용 양액배지인 Sonneveld 양액을 이용하여 양분결핍에 의해 왜성을 나타내는 토마토를 선발하였다. 선발된 토마토에 대해 이차원 전기영동법으로 단백질체를 분석한 결과, 발현차를 나타내는 28개의 단백질 spot이 분리되었다. 분리된 단백질 spot중 현저하게 발현이 억제된 단백질 spot 6개를 선발하여 단백질 서열을 결정하였다. 실험 결과, pI 4.5, 분자량 24 kDa를 나타내는 단백질은 브라시노스테로이드 생합성에 관여하며 왜성 표현형을 나타내는 신호전달 단백질, BAK1으로 동정되었다. BCK1, cystein proteinase, sulfutase, peroxidase, zinc finger factor로 동정 된 나머지 단백질들은 브레시노스테로이드 생합성관련 신호전달기작에 관여하는 단백질로 추정되었다. BAK1을 검정하기 위해 단백질 서열이 결정된 부위로부터 프라이머를 디자인하여 RT-PCR를 수행한 결과, 증폭된 500 bp의 산물이 정상과 발현차를 보여주었는데 이 결과는 양분조절에 의해서도 BAK1의 발현이 조절될 수 있음을 시사한다.

Keywords

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