Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Construction and Characterization of Vector Expressing Low Level of Translation Factor eIF5B

단백질합성인자 eIF5B의 저 발현 효모벡터의 제조 및 특성

  • Published : 2004.03.01
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Abstract

eIF5B is a translation initiation factor that delivers Met-$tRNA^{Met}$ to AUG start codon and subsequently joins the small and large ribosomes. In order to study the function of eIF5B encoded by FUN12, we constructed FUN12 which lacked 5' end of its sequence. We found that this construct lacking almost all of its promoter in pRS plasmid partially complemented slow growth phenotype of fun12 deletion strain. Interestingly, this construct expressed N-terminally truncated eIF5B and its expression level was about 5% of that of wild type eIF5B. Low amount of the eIF5B expressed additionally in fun12 deletion strain played a direct role as a limiting factor for its growth. This limiting factor eIF5B in those strains also modulates activities of overall translation in vitro.

eIF5B는 단백질합성의 개시 인자로서 Met-$tRNA^{Met}$을 AUG 개시코돈에 전달하고, 리보솜의 두 소단위체 결합을 유도한다. 이 인자의 기능을 연구할 목적으로 eIF5B를 코딩하는 FUN12 유전자의 5'말단 부위를 삭제하는 연구를 수행하였다. 프로모터의 대부분을 삭제한 FUN12를 함유한 pRS효모벡터를 FUN12가 삭제되어 천천히 자라는 돌연변이주 ($fun12{\Delta}g$)에 전달하였을 때 그 표현형을 부분적으로 상보하였다. 위와 같이 제조된 벡터에서 N-말단이 상실된 eIF5B 단백질이 발현되었고, 그 양이 정상 균주에서 발현되는 eIF5B 양의 약 5%에 불과하였다. 이와 같이 부분적으로 성장을 상보한 균주에서 발현된 적은 양의 단백질합성개시 인자 eIF5B는 직접적으로 그 성장을 제한하는 요소로 작용한다. 이러한 균주에서 성장의 제한인자인 eIF5B는 in vitro 에서도 역시 전체 단백질 합성의 활성을 조절하였다.

Keywords

References

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