Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Inactivation of the DevS Histidine Kinase of Mycobacterium smegmatis by the Formation of the Intersubunit Disulfide Bond

Subunit 간의 disulfide 결합 형성에 의한 Mycobacterium smegmatis DevS histidine kinase의 불활성화

  • Lee, Jin-Mok (Department of Microbiology, Pusan National University) ;
  • Park, Kwang-Jin (Department of Microbiology, Pusan National University) ;
  • Kim, Min-Ju (Department of Microbiology, Pusan National University) ;
  • Ko, In-Jeong (Korea Science Academy of KAIST) ;
  • Oh, Jeong-Il (Department of Microbiology, Pusan National University)
  • 이진목 (부산대학교 미생물학과) ;
  • 박광진 (부산대학교 미생물학과) ;
  • 김민주 (부산대학교 미생물학과) ;
  • 고인정 (KAIST 부설 한국영재학교) ;
  • 오정일 (부산대학교 미생물학과)
  • Received : 2010.06.04
  • Accepted : 2010.06.16
  • Published : 2010.06.30
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Abstract

The DevSR two-component system is a major regulatory system involved in redox sensing in Mycobacterium smegmatis. The DevSR system consists of the DevS histidine kinase and its cognate DevR response regulator. When exposed to hypoxic conditions, the DevS histidine kinase is activated to phosphorylate the DevR response regulator, leading to the transcriptional activation of the DevR regulation. The ligand-binding state of the heme embedded in the N-terminal GAF domain of DevS determines the kinase activity of DevS. In this study, we demonstrated that the redox-responsive cysteine (C547) in the C-terminal kinase domain is involved in the redox-dependent control of DevS kinase activity. The formation of an intersubunit disulfide bond between the C547 residues in the presence of $O_2$ led to inactivation of DevS kinase activity. The reduction of the oxidized DevS with reductants such as $\beta$-mercaptoethanol and dithiothreitol resulted in the restoration of DevS kinase activity. It was demonstrated in vivo by complementation test that the substitution of C547 to alanine partially impaired the sensory function of DevS in M. smegmatis.

DevSR two-component system은 Mycobacterium smegmatis의 redox sensing에 관련된 주요한 regulatory system이다. DevSR system은 DevS histidine kinase와 DevR response regulator로 구성되어 있다. 저산소 조건에서 DevS histidine kinase는 활성화되어 DevR response regulator를 인산화 시키고, 인산화된 DevR response regulator는 DevR regulon의 transcriptional activator로 작용한다. DevS의 kinase activity는 DevS의 N-terminal에 위치한 GAF domain에 존재하는 heme의 ligand-binding state에 의해 결정된다. 본 연구에서는 C-terminal kinase domain의 redox-responsive cysteine (C547)이 DevS kinase activity의 redox-dependent control과 연관이 있음을 밝혔다. 산소가 존재할 때, C547 residue 사이의 disulfide bond의 형성은 DevS kinase activity를 불활성화 시킨다. $\beta$-mercaptoethanol과 dithiothreitol과 같은 환원제를 이용하여 산화된 DevS를 환원시켰을 때, DevS kinase activity가 복원된 것이 관찰되었다. 또한, C547을 alanine으로 치환했을 때, M. smegmatis의 DevS의 sensory 기능을 부분적으로 손상되는 것이 complementation 실험을 통해 in vivo 상에서 증명되었다.

Keywords

References

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