Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Identification and Characterization of External Copper Responsive Genes of Deinococcus radiodurans

DNA Microarry를 이용한 Deinococcus radiodurans의 구리이온 특이 반응 유전자 탐색 및 특성 분석

  • Joe, Min-Ho (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Lim, Sang-Yong (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Jung, Sun-Wook (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Song, Du-Sub (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Choi, Young-Ji (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Kim, Dong-Ho (Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute)
  • 조민호 (한국원자력연구원 방사선생명공학연구부) ;
  • 임상용 (한국원자력연구원 방사선생명공학연구부) ;
  • 정선욱 (한국원자력연구원 방사선생명공학연구부) ;
  • 송두섭 (한국원자력연구원 방사선생명공학연구부) ;
  • 최영지 (한국원자력연구원 방사선생명공학연구부) ;
  • 김동호 (한국원자력연구원 방사선생명공학연구부)
  • Published : 2008.09.30
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Abstract

Global gene expression of Deinococcus radiodurans, a highly radiation resistant bacterium, in response to excess copper was analyzed by using oligonucleotide microarray chip. Among 3,187 open reading frames of D. radiodurans, seventy genes showed a statistically significant expression ratio of at least 2-fold changes under growth conditions of excess copper; 64 genes were induced and 6 genes were reduced. Especially, two operons ($DRB0014{\sim}DRB0017$ and $DRB0125{\sim}DRB0121$) presumably involved in the iron transport and utilization were the most highly induced genes by excess copper. A quantitative real-time PCR assay revealed that DRB00l4 and DRB0125 are highly transcribed responding to excess copper and 2,2'-dipyridyl, an iron chelator. In addition, the transcription of both genes was not changed by excess iron and bathocuproine disulphonate, a copper chelator. These results suggested that the copper metabolism may be closely connected with the iron transport and utilization in D. radiodurans. However, the disruption of each gene, DRB00l4 and DRB0125, did not affect the copper and radiation resistance, the most well-known character of this organism.

대표적인 방사선저항성 미생물인 Deinococcus radiodurans의 구리이온($CuCl_2,250{\mu}M$)에 대한 발현체 분석을 DNA microarray를 이용하여 수행하였다. 총 3,187개의 open reading frame중 70개의 유전자 발현이 2배 이상증가(64개) 또는 감소(6개)하였다. 이들 중 흥미롭게도 철이온 흡수 관련 유전자들로 추정되는 두 개 operon ($DRB0014{\sim}DRB0017$과 $DRB0121{\sim}DRB0125$)의 발현이 가장 높게 증가하였다. 두 operon의 첫 번째 유전자인 DRB0014와DRB0125의 발현을 실시간 정략 PCR을 이용하여 분석한 결과, 두 유전자 모두 구리 이온($CuCl_2,250{\mu}M$) 또는 철이온 킬레이트화합물(2,2'-dipyridyl, $ 250{\mu}M$)을 첨가하였을 경우 발현이 10배 이상 증가하였으나 철이온($FeCl_3,250{\mu}M$) 또는 구리 이온 킬레이트화합물(bathocuproine disulphonate, $250{\mu}M$)이 존재할 때에는 발현이 변화하지 않았다. 따라서 D. radiodurans의 구리 대사는 철 흡수 시스템과 관련이 있는 것으로 추정된다. 그러나 DRB0014와DRB0125의 변이는D. radiodurans의 구리 저항성 및 방사선 저항성에 큰 영향을 미치지 않는 것으로 관찰되었다.

Keywords

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