생명과학회지 (Journal of Life Science)

  • 제19권11호
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  • Pages.1506-1513
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  • 2009
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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캐너비노이드 수용체 CB2의 신호전달작용에 미치는 RGS3의 억제적 효과

RGS3 Suppresses cAMP Response Element (CRE) Activity Mediated by CB2 Cannabinoid Receptor in HEK293 Cells

  • 김성대 (경북대학교 수의과대학 수의학과) ;
  • 이휘민 (경북대학교 수의과대학 수의학과) ;
  • 메하리 엔델 (경북대학교 수의과대학 수의학과) ;
  • 조재열 (강원대학교 생명공학부 분자의생명공학) ;
  • 박화진 (인제대학교 의생명공학대학 임상병리학과) ;
  • 오재욱 (건국대학교 동물생명과학대학 축산식품생물공학) ;
  • 이만휘 (경북대학교 수의과대학 수의학과)
  • Kim, Sung-Dae (Laboratory of Physiology & Signaling, College of Veterinary Medicine, Kyungpook National University) ;
  • Lee, Whi-Min (Laboratory of Physiology & Signaling, College of Veterinary Medicine, Kyungpook National University) ;
  • Endale, Mehari (Laboratory of Physiology & Signaling, College of Veterinary Medicine, Kyungpook National University) ;
  • Cho, Jae-Youl (School of Bioscience and Biotechnology, Kangwon National University) ;
  • Park, Hwa-Jin (College of Biomedical Science and Engineering, Inje University) ;
  • Oh, Jae-Wook (College of Animal Bioscience & Technology, Konkuk University) ;
  • Rhee, Man-Hee (Laboratory of Physiology & Signaling, College of Veterinary Medicine, Kyungpook National University)
  • 발행 : 2009.11.30
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초록

RGS단백질은 G 단백질 신호전달작용에 있어서 신호를 억제하는 조절단백질로서 G 단백질 매개수용체(GPCR)의 활성을 억제하는 것으로 알려졌다. 그렇지만 캐너비노이드 수용체 CB2의 활성에 있어서 RGS 단백질의 조절효과에 관해서는 지금까지 알려져 있지 않다. 그러므로 본 연구에서 우리는 RGS2, 3, 4, 5와 캐너비노이드 수용체 CB2 cDNA를 동시에 HEK293 세포주에 발현시킨 후 각 RGS 단백질의 효과를 조사하였다. CB2 단백질을 발현하는 HEK293 세포주(CB2-HEK293)에서 CB2 효현제인 WIN55,212-2는 폴스콜린으로 유도된 cAMP response element (CRE) 활성을 억제하였다. 이러한 WIN55,212-2의 CRE 억제 활성은 RGS3에 의하여 차단되었지만 RGS2, 4, 및 RGS5에서는 관찰되지 않았다. 뿐만 아니라 RGS3 small interference RNA (siRNA)를 사용하여 내인성 RGS3 단백질의 발현을 저하시키면 WIN55,212-2에 의한 폴스콜린 유도 CRE 억제활성은 더욱 증강되었다. 이상의 결과는 캐너비노이드 수용체 CB2 신호전달작용에 있어서 RGS 단백질의 기능적 역할과 특히 내인성 RGS3의 캐너비노이드 수용체 CB2에 대한 선택적 작용을 나타낸다.

RGS proteins have been identified as negative regulators of G protein signalling pathways and attenuate the activity of GPCR receptors. However, information on the regulatory effects of RGS proteins in the activity of cannabinoid receptors is limited. In this study, the role of RGS proteins on the signal transduction of the CB2 cannabinoid receptor was investigated in HEK293 cells co-transfected with CB2-receptors and plasmids encoding RGS2, RGS3, RGS4 and RGS5. Treatment of cells with WIN55, 212-2, a CB2 receptor agonist, inhibited forskolin-induced cAMP response element (CRE) activity in CB2-transfected HEK293 (CB2-HEK293) cells. This inhibitory effect of WIN 55, 212-2 on CRE activity was reversed by co-transfection of CB2-HEK293 cells with RGS3, but not with RGS2, RGS4 and RGS5. However, endogenous RGS3 protein knocked down by a small interfering siRNA targeting RGS3 gene enhanced inhibition of forskolin induced CRE activity via agonist induced CB2 receptor signal transduction. These results indicate the functional role of endogenous RGS protein in cannabinoid signaling pathways and define receptor-selective roles of endogenous RGS3 in modulating CRE transcriptional responses to agonist induced CB2 receptor activity.

키워드

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