Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Construction of spDbp5 Null Mutants Defective in mRNA Export

분열효모에서 spDbp5 유전자의 결실돌연변이 제조와 기능에 대한 연구

  • Bae, Jin-Ah (Basic Science Research Institute and Department of Biology, College of Natural Sciences, Sungshin Women's University) ;
  • Cho, Hyun-Jin (Basic Science Research Institute and Department of Biology, College of Natural Sciences, Sungshin Women's University) ;
  • Yoon, Jin-Ho (Basic Science Research Institute and Department of Biology, College of Natural Sciences, Sungshin Women's University)
  • 배진아 (성신여자대학교 자연과학대학 생물학과 및 기초과학연구소) ;
  • 조현진 (성신여자대학교 자연과학대학 생물학과 및 기초과학연구소) ;
  • 윤진호 (성신여자대학교 자연과학대학 생물학과 및 기초과학연구소)
  • Published : 2008.03.31
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Abstract

We constructed the null mutants of fission yeast Schizosaccharomyces pombe spDbp5 gene that is homologous to DEAD-box RNA helicase DBP5 in budding yeast Saccharomyces cerevisiae, which plays important roles in mRNA export out of nucleus. A null mutant in an $h^+/h^+$ diploid strain was constructed by replacing the spDbp5-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spDbp5 is essential for vegetative growth. The haploid spDbp5 null mutants harboring pREP81X-spDbp5 plasmid showed extensive $poly(A)^+$ RNA accumulation in the nucleus and decrease in the cytoplasm after repression of spDbp5 expression. These results suggest that spDbp5 is also involved in mRNA export from the nucleus.

mRNA의 핵에서 세포질로의 이동에 중요한 역할을 하는 발아효모 Saccharomyces cerevisiae의 DEAD-box RNA helicase인 DBP5 유전자와 유사한 분열효모 Schizosaccharomyces pombe의 유전자(spDbp5로 명명)의 결실돌연변이주(knockout mutant)를 제조하여 그 특성을 조사하였다. 이배체인 S. pombe 균주에 하나의 spDbp5 유전자만을 결실시킨 후 4분체분석(tetrad analysis)을 수행한 결과, 이 유전자가 결실된 반수체 균주는 생장하지 못했다. mRNA의 핵에서 세포질로의 이동에 있어서 spDbp5의 역할을 알아보기 위해, spDbp5의 발현이 티아민(thiamin)에 의해 억제되는 균주를 제작하여 in situ hybridization을 통해 세포 내의 $poly(A)^+$ RNA 분포를 살펴보았다. spDbp5 유전자의 발현이 억제되면, $poly(A)^+$ RNA가 핵 안에 축적되고세포질에서는 줄어들었다. 이와 같은 결과들은 spDbp5 유전자 역시 mRNA의 핵에서 세포질로의 이동에 매우 중요한 역할을 담당하고 있음을 시사한다.

Keywords

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