A. c-Met harbors a putative leucine-rich nuclear export signal (LR-NES)
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73 - 국문요약 -
c-Met C-말단 조각의 핵-세포질 운반에 관여하는 단백질 부위:
새로운 "pH-의존성" nuclear localization signal
아주대학교 대학원 의생명과학과 (종양생물학 전공)
Shubhash Chandra Chaudhary
(지도교수 : 이재호)
c-Met 수용체 티로신 카이나제의 C-말단 조각은 간세포성장인자의 유무와 무관하게 여러 세포에서 관찰된 바 있다. 하지만 이 단백질 조각이 어떻게 핵 안으로 이동되는지에 대해서는 잘 알려져 있지 않다. 본 저자는 c-Met 의 세포막인접부위에 존재하는 두개의 히스티딘 잔기 포함 부위(H1068-H1079)가 새로운 NLS 후보임을 밝혔다. 세포면역염색과 세포성분분리 방법으로 살펴 본 결과, 이 부위를 결손시키면 HeLa 세포에서 c-Met C-말단부위의 핵내 이동이 뚜렷하게 억제됨을 확인할 수 있었다. 또한 두개의 히스티딘 잔기를 알라닌으로 치환해도 같은 결과를 얻을 수 있었다. 히스티딘 잔기의 치환에 의한 핵내 이동 감소현상은 통상적으로 알려진 핵내 이동의 운반체인 importin β 와 c-Met 조각의 결합 감소를 동반하였다. c-Met 에서 발견된 이 NLS 부위는 히스티딘 잔기를 포함하고 있다는 점에서 독특하다고 할 수 있다. 히스티딘은 라이신이나 아르기닌과 달리 전기적으로 양성을 띄는 정도가 pH 에 의해서 영향을 받는다. 특히 pH 6~7
74
범위에서 그러한 현상이 명확이 나타난다. 본 저자는 이에 착안하여 이 NLS 가 pH 의 변화에 따라 그 활성이 달라지는지 확인했다. 이를 위해 Na+/H+ exchanger 인 nigericin 과
pH6.5 인 KRB 완충액을 사용하여 세포질 내 pH 를 낮추었을 때, 예상대로 핵내로 더 많은 c-Met 조각이 이동함을 확인할 수 있었고 importin β 와 c-Met 조각 간의 결합도 증가함을 확인할 수 있었다. 더 나아가 nigericin 처리에 의해 HeLa 세포에 본래 존재하던
c-Met C-말단 조각의 핵내 이동이 역시 증가함을 확인할 수 있었다. 한편 히스티딘 잔기를 라이신이나 아르기닌으로 치환하자 "pH-의존성"이 소실됨을 확인할 수 있었다.
본 저자가 알고 있는 한, c-Met 에서 발견된 NLS 는 최초로 발견된 "pH-의존성" NLS 이다.
본 저자는 여기에 더해 c-Met 의 세포막인접부위에 고전적으로 알려져 있는 nuclear export
signal 이 존재함을 확인할 수 있었다. c-Met 의 경우 세포막인접부위에 nuclear localization signal 과 nuclear export signal 이 함께 존재함으로써 C-말단 조각의 핵내외 위치를 조절함을 알 수 있었다.
핵심되는 말 ; c-Met, 암유전자, 수용체 티로신 키나아제, 간세포성장인자, nuclear
localization signal, nuclear export signal, 세포막인접부위