Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
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Cloning of hnRNP E1 cDNA via yeast two-hybrid system and a study on protein-protein interaction between hnRNP E1 and hnRNP K

이스트 two-hybrid 시스템을 이용한 hnRNP E1 cDNA의 클로닝과 hnRNP E1-hnRNP K 상호결합에 대한 연구

  • 최미영 (선문대학교 건강과학대학 의생명과학과)
  • Published : 2008.12.31
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Abstract

The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a component of hnRNP complexes. This protein binds strongly to cytidine-rich RNA/DNA sequences. It is a nucleocytoplasmic shuttling protein. To investigate the functions of hnRNP K, I searched for hnRNP K-interacting proteins in HeLa cDNA library using a yeast two-hybrid screening system. One of the cDNA clones is identical to human hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585). In this study, hnRNP K is shown to specifically interact with hnRNP E1 in yeast two-hybrid system and in vitro biochemical assay.

hnRNP K 단백질은 hnRNP 복합체를 구성하는 핵단백질들 중의 하나이며 시토신이 많은 RNA/DNA sequence에 잘 결합한다. 이 단백질은 핵 내에서만 머무르지 않고 핵과 세포질을 왕복하는 특징을 지니고 있다. hnRNP K의 기능을 조사하기 위하여 우선 hnRNP K와 상호 결합하는 세포내 단백질을 찾아내고자 하였다. 이를 위하여 본 연구에서는 이스트 two-hybrid 시스템을 사용하여 HeLa CDNA librar를 탐색하였다. 그 결과 얻은 클론들 중에는 사람의 hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585) 클론이 포함되어 있었다. 본 논문에서는 이스트 two-hybrid 시스템과 in vitro에서의 생화학적 실험을 통하여 hnRNP E1은 hnRNP K와 특이적으로 상호 결합한다는 것을 밝혔다.

Keywords

References

  1. G. Dreyfuss, M. J. Matunis, S. Pinol-Roma, and C. G. Burd, "hnRNP proteins and the biogenesis of mRNA". Annu. Rev. Bioche, Vol.. 62, pp. 289-321. 1993. 12. https://doi.org/10.1146/annurev.bi.62.070193.001445
  2. A. M. Krecic and M. S. Swanson "hnRNP complexes: composition, structure, and function", Curr. Opin. Cell Biol. Vol. 11, pp. 363-371. 1999. 10. https://doi.org/10.1016/S0955-0674(99)80051-9
  3. M. J. Matunis, W. M. Michael and G. Dreyfuss "Characterization and primary structure of the poly (C)-binding heterogeneous nuclear ribonucleoprotein complex K protein", Mol. Biol. Cell, Vol. 12, pp. 164-171. 1992. 6. https://doi.org/10.1128/MCB.12.1.164
  4. H. Siomi, M. Choi, M. C. Siomi, R. L. Nussbaum, and G. Dreyfuss "Essential role for KH domains in RNA-binding: impaired RNA binding by a mutation in the KH domain of FMR1 that causes fragile X syndrome", Cell, Vol. 77, pp. 33-39. 1994. 4. https://doi.org/10.1016/0092-8674(94)90232-1
  5. K. Bomsztyk, O. Denisenko and J. Ostrowski J "hnRNP K: one protein multiple processes", BioEssays, Vol. 26, pp. 629-638. 2004. 6. https://doi.org/10.1002/bies.20048
  6. H. E. Feilotter, G. J. Hannon, C. J. Ruddell, and D. Beach "Construction of an improved host strain for two hybrid screening", Nucl. Acids Res, Vol.. 22, pp. 1502-1503. 1994. 8. https://doi.org/10.1093/nar/22.8.1502
  7. B. Hahm, O. H. Cho, J. E. Kim, Y. K. Kim, J. H. Kim, Y. L. Oh, and S. K. Jang "Polypyrimidine tract-binding protein interacts with HnRNP L", FEBS Letters, Vol. 425, pp. 401-406. 1998. 10. https://doi.org/10.1016/S0014-5793(98)00269-5
  8. J. H. Kim, B. Hahm, Y. K. Kim, M. Choi, and S. K. Jang, "Protin-Protein Interaction Among hnRNPs Shuttling Between Nucleus and Cytoplasm" J. Mol. Biol. Vol. 298, pp. 395-405. 2000. 6. https://doi.org/10.1006/jmbi.2000.3687
  9. J-Y. Yoon and M. Choi. "Heterogeneous nuclear ribonucleoprotein D-like protein JKTBP interacts with heterogeneous nuclear ribonucleoprotein K", Genes and Genomics, Vol. 30, pp. 419-426. 2008. 8.
  10. M. Choi. "hnRNP E1 interacts with hnRNP L", Genes and Genomics, Vol. 30, pp. 355-363. 2008. 8.
  11. M. Kiledjian, X. Wang and S. A. Liebharber, "Identification of two KH domain prteins in the ${\alpha}-globin$ mRNP stability complex", EMBO J. Vol. 14, pp. 4357-4364. 1995. 6.
  12. B-J. Thiele, A. Doller, T. Kahne, R. Pregla, R. Hetzer, and V. Regitz-Zagrosek, "RNA-binding proteins heterogeneous nuclear ribonucleoprotein A1, E1, and K are involved in post-transcriptional control of collagen I and III synthesis", Circ. Res. Vol. 95, pp. 1058-1066. 2004. 10. https://doi.org/10.1161/01.RES.0000149166.33833.08