Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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A putative prolyl tRNA synthetase is involved in pheromone induction in Schizosaccharomyces pombe

Schizosaccharomyces pombe의 pheromone 유도와 연관된 prolyl tRNA synthetase

  • Kim, Daemyung (Department of Biomedical Science, Cheongju University)
  • 김대명 (청주대학교 바이오메디컬학과)
  • Received : 2018.09.14
  • Accepted : 2018.10.03
  • Published : 2018.12.31
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Abstract

Previously, six Schizosaccharomyce pombe mutants that induce pheromone even in the presence of nitrogen source were isolated from a bank of temperature sensitive mutants. In this report, one of these mutants, pws6 was further characterized. The pheromone induction in pws6 mutant cells was specific to nutrient: the M-factor pheromone was induced without nitrogen starvation but not without glucose starvation. This result suggests that the pws6 mutant might have a specific defect in the pathway for nitrogen starvation. The pws6 mutant induces P-factor pheromone as well as M-factor without starvation of nitrogen in temperature sensitive mode, suggesting that the pheromone induction phenotype of pws6 mutation is not cell-type specific. From cloning of the $pws6^+$ gene by complementation of the temperature sensitive growth defect, three plasmids containing 8.1 kb, 3.3 kb, and 4.8 kb yeast DNA were recovered. These plasmids complement the growth defect of the pws6 mutant by 100%, 70%, and 10~20%, respectively. The abilities of these plasmids to complement pheromone induction phenotype of pws6 mutant cells were correlated well with the efficiencies of complementation of the growth defect. With comparison of their open reading frames to the complementation efficiencies, it is concluded that the open reading frame, SPBC19C7.06 is responsible for the complementation of temperature sensitive phenotype of the pws6 mutant. This open reading frame, named prs1, contains one long exon with no intron and encodes a putative prolyl tRNA synthetase. The putative Prs1 protein exhibits significant similarities to the prolyl tRNA synthetases of other species.

이전의 연구에서 질소원이 존재하여도 페로몬을 유도하는 6개의 Schizosaccharomyces pombe 돌연변이체를 온도민감성 돌연변이체들의 저장고로부터 분리하였음이 보고된 바 있다. 본 연구에서는 이들 중 하나인 pws6 돌연변이체의 특성을 더 연구하였다. 이 돌연변이체는 영양물질에 특이적으로 페로몬 유도를 나타내었다. 즉 질소의 고갈은 없어도 M-factor 페로몬을 유도하였으나 탄소의 고갈이 없으면 유도되지 않았다. 이러한 결과는 pws6 돌연변이체가 질소 고갈을 전달하는 경로에 특이한 결함을 가지고 있음을 시사한다. 이 돌연변이체는 M-factor 페로몬뿐만 아니라 P-factor 페로몬도 온도에 민감한 양식으로 질소의 고갈 없이 유도함을 보여 주어 이 돌연변이체의 페로몬 유도는 세포 유형에 특이적이지 않음을 시사하였다. 이 돌연변이체의 온도 민감성 성장 결함의 상보적 보완에 의해 $pws6^+$ 유전자를 클로닝하여 8.1 kb, 3.3 kb, 그리고 4.8 kb 효모 DNA를 가진 3개의 플라스미드가 분리되었다. 이 플라스미드들은 pws6 돌연변이체의 성장 결함을 각각 100%, 70%, 그리고 10-20% 보완하였다. 또한 이 플라스미드들은 pws6 돌연변이체의 페로몬 유도 특성을 보완하는 능력을 가지고 있었으며 이는 돌연변이체의 성장 결함 보완 효율과 밀접한 연관성이 있음을 보여 주었다. 이들의 오픈 리딩 프레임을 성장 결함의 보완 효율과 비교하여 오픈 리딩 프레임 SPBC19C7.06이 pws6 돌연변이체의 온도 민감성 특성을 상보적으로 보완하는데 원인이 되는 리딩 프레임으로 결론 내렸다. 이 오픈 리딩 프레임은 prs1으로 명명되었으며 인트론이 없이 하나의 긴 엑손을 가지고 있는 추정된 prolyl tRNA synthetase를 암호화한다. 추정된 Prs1 단백질은 다른 종의 prolyl tRNA synthetase와 상당한 유사성을 보여 주었다.

Keywords

MSMHBQ_2018_v54n4_309_f0001.png 이미지

Fig. 1. Effect of nutrient on the M-factor pheromone induction in pws6 mutant cells.

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Fig. 2. P-factor pheromone induction in pws6 mutant cells.

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Fig. 3. Restriction map and complementation analysis of the pws6 clones.

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Fig. 4. Complementation of pheromone induction phenotype.

Table 1. Transformation of the pws6 cells with genomic DNA

MSMHBQ_2018_v54n4_309_t0001.png 이미지

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