생명과학회지 (Journal of Life Science)

  • 제27권10호
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  • Pages.1191-1198
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  • 2017
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

Ras-GTPase-activating protein SH3 domain-binding proteins 2, G3BP2와 KIF5A C-말단 꼬리 영역과의 결합

Interaction of Ras-GTPase-activating Protein SH3 Domain-binding Proteins 2, G3BP2, With the C-terminal Tail Region of KIF5A

  • 정영주 (인제대학교 의과대학 생화학교실) ;
  • 장원희 (인제대학교 의과대학 생화학교실) ;
  • 이원희 (인제대학교 의과대학 신경외과학교실) ;
  • 김무성 (인제대학교 의과대학 신경외과학교실) ;
  • 김상진 (인제대학교 의과대학 신경과학교실) ;
  • 엄상화 (인제대학교 의과대학 예방의학교실) ;
  • 문일수 (동국대학교 의과대학 해부학교실) ;
  • 석대현 (인제대학교 의과대학 생화학교실)
  • Jeong, Young Joo (Department of Biochemistry, Inje University College of Medicine) ;
  • Jang, Won Hee (Department of Biochemistry, Inje University College of Medicine) ;
  • Lee, Won Hee (Department of Neurosurgery, Inje University College of Medicine) ;
  • Kim, Mooseong (Department of Neurosurgery, Inje University College of Medicine) ;
  • Kim, Sang-Jin (Department of Neurology, Inje University College of Medicine) ;
  • Urm, Sang-Hwa (Department of Preventive Medicine, Inje University College of Medicine) ;
  • Moon, Il Soo (Departments of Anatomy, College of Medicine, Dongguk University) ;
  • Seog, Dae-Hyun (Department of Biochemistry, Inje University College of Medicine)
  • 투고 : 2017.07.07
  • 심사 : 2017.09.07
  • 발행 : 2017.10.30
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초록

세포 내 소기관들과 소포들은 세포 내에서 미세소관을 따라 적절한 구획으로 수송된다. 이러한 세포 내 수송과정은 분자 모터단백질인 kinesin과 dynein에 의하여 이루어진다. Kinesin 1은 오징어 축삭돌기 세포질로부터 처음 분리되었으며 2개의 중쇄단위체(KHCs, 또는 KIF5s) 및 이와 결합하는 경쇄단위체(KLCs)의 복합체를 형성한다. KIF5s는C-말단 고리 영역을 통해 많은 다양한 단백질과 결합하는데, 아직 그 결합단백질들은 충분히 밝혀지지 않았다. 본 연구에서는 KIF5A 결합단백질을 분리하기 위하여 효모 two-hybrid 탐색을 수행하여 스트레스 과립형성과 mRNP 위치결정에 관여하는 Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2)를 분리하였다. G3BP2는 KIF5A의 C-말단 고리 영역에 존재하는 73개 아미노산을 포함하는 영역과 결합하였다. 그러나 G3BP2는 KIF5B, KIF5C, KLC1, KIF3A와는 결합하지 않았다. KIF5A는 G3BP2의 arginine-glycine-glycine(RGG)/Gly-rich 도메인과 결합하지만 G3BP1과는 결합하지 않았다. HEK-293T세포에 G3BP2와 KIF5A를 발현하여 면역침강한 결과 G3BP2와 KIF5A는 같이 침강하였다. 또한 HEK-293T 세포 내의 전체에서 두 단백질은 같은 부위에 존재하였다. 이러한 결과들은 세포 내에서 G3BP2는 KIF5A와 결합하는 결합단백질로 확인 되었다.

Vesicles and organelles are transported along microtubule and delivered to appropriate compartments in cells. The intracellular transport process is mediated by molecular motor proteins, kinesin, and dynein. Kinesin is a plus-end-directed molecular motor protein that moves the various cargoes along microtubule tracks. Kinesin 1 is first isolated from squid axoplasm is a dimer of two heavy chains (KHCs, also called KIF5s), each of which is associated with the light chain (KLC). KIF5s interact with many different binding proteins through their carboxyl (C)-terminal tail region, but their binding proteins have yet to be specified. To identify the interacting proteins for KIF5A, we performed the yeast two-hybrid screening and found a specific interaction with Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2), which is involved in stress granule formation and mRNA-protein (mRNP) localization. G3BP2 bound to the C-terminal 73 amino acids of KIF5A but did not interact with the KIF5B, nor the KIF5C in the yeast two-hybrid assay. The arginine-glycine-glycine (RGG)/Gly-rich region domain of G3BP2 is a minimal binding domain for interaction with KIF5A. However, G3BP1 did not interact with KIF5A. When co-expressed in HEK-293T cells, G3BP2 co-localized with KIF5A and was co-immunoprecipitated with KIF5A. These results indicate that G3BP2, which was originally identified as a Ras-GAP SH3 domain-binding protein, is a protein that interacts with KIF5A.

키워드

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