Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Defensive roles of Sdu1, a PPPDE superfamily member with ubiquitin C-terminal hydrolase activity, against thermal stress in Schizosaccharomyces pombe

카르복시 말단 유비퀴틴 가수분해 효소 활성 보유 PPPDE superfamily member인 Schizosaccharomyces pombe Sdu1의 열 스트레스에 대한 방어적 역할

  • Han, Hee (Department of Biochemistry, Kangwon National University) ;
  • Heo, Tae Young (Department of Biochemistry, Kangwon National University) ;
  • Ryu, In Wang (Department of Biological Sciences, Kangwon National University) ;
  • Kim, Kyunghoon (Department of Biological Sciences, Kangwon National University) ;
  • Lim, Chang-Jin (Department of Biochemistry, Kangwon National University)
  • 한희 (강원대학교 자연과학대학 생화학과) ;
  • 허태영 (강원대학교 자연과학대학 생화학과) ;
  • 류인왕 (강원대학교 자연과학대학 생명과학과) ;
  • 김경훈 (강원대학교 자연과학대학 생명과학과) ;
  • 임창진 (강원대학교 자연과학대학 생화학과)
  • Received : 2015.08.31
  • Accepted : 2015.11.04
  • Published : 2015.12.31
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Abstract

The $sdu1^+$ gene encodes Sdu1, a PPPDE superfamily member of deubiquitinating enzymes (DUBs) in Schizosaccharomyces pombe. Sdu1 was previously shown to contain an actual ubiquitin C-terminal hydrolase (UCH) activity using the recombinant plasmid pYSTP which harbors the $sdu1^+$ gene. This work was designed to assess a thermotolerant role of Sdu1 against high incubation temperatures. In the temperature-shift experiments, the S. pombe cells harboring pYSTP grew much better after the shifts to $37^{\circ}C$ and $42^{\circ}C$, when compared with the vector control cells. After being shifted to $37^{\circ}C$ and $42^{\circ}C$ for 6 h, the S. pombe cells harboring pYSTP contained lower reactive oxygen species (ROS) levels, compared with the vector control cells. The nitric oxide (NO) levels of the S. pombe cells harboring pYSTP were slightly lower than those of the vector control cells in the absence or presence of the temperature shifting. The total glutathione (GSH) levels of the S. pombe cells harboring pYSTP were significantly higher than those of the vector control cells. Total superoxide dismutase (SOD) and GSH peroxidase activities were also higher in the S. pombe cells harboring pYSTP after the temperature shifts than in the vector control cells. In brief, the S. pombe Sdu1 plays a thermotolerant role against high incubation temperature through the down-regulation of ROS and NO and the up-regulation of total GSH content, total SOD and GSH peroxidase activities.

Schizosaccharomyces pombe $sdu1^+$ 유전자는 PPPDE superfamily member에 속하는 탈유비퀴틴화 효소인 Sdu1을 엔코딩한다. 이전 연구에서, $sdu1^+$ 유전자 포함 재조합 플라스미드 pYSTP를 사용하여 Sdu1이 카르복시 말단 유비퀴틴 가수분해 활성을 보유한다는 사실이 입증된 바 있다. 본 연구에서는, 높은 배양 온도에 대한 Sdu1의 열내성적 역할이 검토되었다. 온도 천이 실험에서, pYSTP 포함 S. pombe 세포들이 37도나 42도로 천이한 후에 벡터 대조 세포들보다 훨씬 더 잘 성장하였다. 37도나 42도로 천이 한 후 6시간 배양한 pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 낮은 활성산소종 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 온도 천이와 관계없이 벡터 대조 세포들보다 다소 낮은 일산화질소 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 훨씬 높은 총 글루타치온 수준을 나타내었다. 온도천이 후의 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 활성들이 pYSTP 포함 S. pombe 세포들에서 더 높은 것으로 측정되었다. 요약하면, S. pombe Sdu1은 활성산소종과 일산화질소 수준은 낮추고, 총 글루타치온, 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 수준들은 증가시킴으로써 높은 배양 온도에 대한 열내성적 역할을 나타낸다.

Keywords

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