Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Reactive oxygen species-dependent down-regulation of ubiquitin C-terminal hydrolase in Schizosaccharomyces pombe

Schizosaccharomyces pombe에서의 유비퀴틴 C-말단 가수분해효소의 활성산소종 의존성 하향조절

  • Jo, Hannah (Department of Biochemistry, College of Natural Sciences, Kangwon National University) ;
  • Lim, Hye-Won (Shebah Biotech Inc.) ;
  • Kwon, Hee-Souk (Hankook Cosmo Co.) ;
  • Lim, Chang-Jin (Department of Biochemistry, College of Natural Sciences, Kangwon National University) ;
  • Park, Kwang Hark (Department of Biological Sciences, College of Natural Sciences, Kangwon National University) ;
  • Jin, Chang Duck (Department of Biological Sciences, College of Natural Sciences, Kangwon National University) ;
  • Kim, Kyunghoon (Department of Biological Sciences, College of Natural Sciences, Kangwon National University)
  • 조한나 (강원대학교 자연과학대학 생화학과) ;
  • 임혜원 (주)세바바이오텍) ;
  • 권희석 (주)한국코스모) ;
  • 임창진 (강원대학교 자연과학대학 생화학과) ;
  • 박광학 (강원대학교 자연과학대학 생명과학과) ;
  • 진창덕 (강원대학교 자연과학대학 생명과학과) ;
  • 김경훈 (강원대학교 자연과학대학 생명과학과)
  • Received : 2016.05.04
  • Accepted : 2016.06.23
  • Published : 2016.06.30
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Abstract

The Schizosaccharomyces pombe $sdu1^+$ gene, belonging to the PPPDE superfamily of deubiquitinating enzyme (DUB) genes, was previously shown to encode a protein with ubiquitin C-terminal hydrolase (UCH) activity and to participate in the response against oxidative and nitrosative stresses. This work focused on the reactive oxygen species (ROS)-dependent regulation of the S. pombe $sdu1^+$ gene. UCH activities, encoded by the $sdu1^+$ gene, were attenuated in the S. pombe cells exposed to $H_2O_2$, superoxide radical-generating menadione (MD), and nitric oxide (NO)-generating sodium nitroprusside (SNP). Reduced glutathione (GSH) and its precursor N-acetylcysteine (NAC) were able to significantly enhance the UCH activities in the absence or presence of $H_2O_2$. However, the influences of both GSH and NAC on the ROS levels in the absence or presence of $H_2O_2$ were opposite to their effects on the UCH activities under the same conditions. The UCH activities in the Sdu1-overexpressing S. pombe cells were also diminished under exposure to $H_2O_2$, MD and SNP, but still remained to be higher than those in the vector control cells. In brief, it is proposed that the S. pombe $sdu1^+$ gene is regulated by ROS in a negative manner, the meaning of which largely remains elusive.

탈유비퀴틴 효소 중 PPPDE 상과에 속하는 Schizosaccharomyces pombe의 $sdu1^+$ 유전자가 유비퀴틴 C-말단 가수분해 효소 활성을 갖는 단백질을 인코딩하고, 산화적 및 일산화질소 스트레스 방어에 관여함이 이전에 밝혀진 바 있다. 예비적인 본 연구는 정상적인 및 과잉발현의 조건에서 S. pombe 유비퀴틴 C-말단 가수분해 효소 활성의 활성산소종 의존성 조절에 초점을 맞추었다. 과산화수소, 수퍼옥사이드 라디칼 생성하는 메나디온 및 일산화질소 생성하는 sodium nitroprusside (SNP)에 노출시킨 S. pombe 세포에서 유비퀴틴 C-말단 가수분해 효소 활성이 감소되었다. 환원형 글루타치온과 그 전구체인 N-acetylcysteine은 과산화수소의 존재 유무에 상관없이 유비퀴틴 C-말단 가수분해 효소 활성을 현저하게 증강시켰다. 그러나, 과산화수소의 부재 시 혹은 존재 시 활성산소종에 미치는 글루타치온과 N-acetylcysteine의 영향은 같은 조건 하에서의 유비퀴틴 C-말단 가수분해 효소 활성 패턴과 상반되었다. 과잉발현의 유비퀴틴 C-말단 가수분해 효소 활성을 보이는 재조합 플라즈미드 pYSTP를 보유하는 S. pombe 세포에서 유비퀴틴 C-말단 가수분해 효소 활성도 과산화수소, 메나디온 및 SNP에의 노출되는 조건에서 감소되었지만, 벡터 대조 세포에서 보다는 높게 유지되었다. 요약하면, S. pombe 유비퀴틴 C-말단 가수분해 효소 활성은 활성산소종에 의하여 하향조절 되지만, 그 의의는 현재로썬 알려지고 있지 않은 상태이다.

Keywords

References

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