Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Identification and Chararterization of Stationary-phase Specific Cytosolic Protein in Salmonella typhimurium

정지기 Salmonella typhimurium 세포에서 특이적으로 발현되는 세포질 단백질의 동정 및 발현조절에 대한 연구

  • Yoo, Ah-Young (Division of Biological Sciences, Pusan National University) ;
  • Kim, Young-Hee (Division of Biological Sciences, Pusan National University) ;
  • Yu, Jong-Earn (Division of Biological Sciences, Pusan National University) ;
  • Kim, Sam-Woong (Division of Biological Sciences, Pusan National University) ;
  • Baik, Hyung-Suk (Division of Biological Sciences, Pusan National University) ;
  • Kang, Ho-Young (Division of Biological Sciences, Pusan National University)
  • 유아영 (부산대학교 생명과학부 미생물학과) ;
  • 김영희 (부산대학교 생명과학부 미생물학과) ;
  • 유종언 (부산대학교 생명과학부 미생물학과) ;
  • 김삼웅 (부산대학교 생명과학부 미생물학과) ;
  • 백형석 (부산대학교 생명과학부 미생물학과) ;
  • 강호영 (부산대학교 생명과학부 미생물학과)
  • Published : 2007.02.28
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Abstract

Salmonella is facultative intracellular pathogen that can survive and replicate in macrophages even though these cells are equipped with a plethora of anti-microbial mechanisms. To survive in this hostile intracellular environment, Salmonella has evolved numerous defense mechartisms. An approximately 20 kDa protein was detected as a stationary-phase specific protein band in cytosolic fraction. It was identified as a DNA binding protein in stationary phase (Dps) by analysis of MALDI-TOF assay. It has been known that Dps, the protein produced in the stationary phase of bacteria, allows DNA to form chromatin by binding to DNA nonspecifically and protects DNA from reactive oxidative species (ROS). For further study, Dps specific polyclonal antibodies were generated by injection of purified Dps protein into rabbit. To examine the Finfluence of several regulatory proteins in the expression dps gene, Dps protein level in various S. typhimurium mutants defecting regulatory proteins were investigated by Westernblot using Dps specific polyclonal antibodies.

Salmoenella는 대표적인 intracellular pathogen으로 숙주의 면역 세포인 macrophage 내에서 살아남아 이들을 매개로 숙주의 몸 전체를 이동해 가면서 전신성 감염을 일으킨다. 살모넬라는 숙주 세포 내부의 이러한 극한 환경을 극복하기 위해서 다양한 방어 기작을 가진다. 본 연구에서는 복합적인 스트레스가 작용하는 정지기 Salmonella에서 특이적으로 발현되는 단백질에 주목하였다. 정지기 상태의 Salmonella에서 약 20 kDa의 단백질이 특이적으로 많이 발현되었으며, 세포질 분획을 통해 이 단백질이 세포질 부분에 존재함을 알 수 있었다. MALDI-TOF 분석을 통해 이 단백질이 $\b{D}NA$ binding $\b{p}rotein$ in $\b{s}tationary$ phase (Dps) 단백질임을 확인하였다. Dps 단백질은 스트레스가 주어진 상황에서 DNA에 비특이적으로 결합하여 DNA가 안정한 형태를 유지하도록 하여 스트레스로부터 염색체를 보호하는 역할을 하는 것으로 알려져 있다. 이후의 연구를 위하여 과발현하여 정제한 Dps 단백질을 토끼에 주사하여 Dps 특이적인 항체를 제조하였다. dps 발현에 영향을 미치는 조절자 단백질을 알기 위하여 다양한 S. typhimrium 돌연변이주들 내에서의 Dps 단백질양을 조사하였다.

Keywords

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