Jong Heon Kim1, Bumsuk Hahm1, Yoon Ki Kim1, Mieyoung Choi2 and Sung Key Jang1,*
1 Department of Life Science Pohang University of Science and Technology, Hyoja-Dong San31 Pohang, Kyungbuk 790-784, South Korea
2 Department of Biological Science, Sunmoon University #100, Kalsan-Ri, Tangjeong-Myeon, Asan, Chungnam 336-840, South Korea
Received 20 December 1999; revised 6 March 2000; accepted 14 March 2000. ; Available online 25 March 2002.
Heterogeneous nuclear ribonucleoproteins (hnRNPs) are involved in several RNA-related biological processes such as transcription, pre-mRNA processing, mature mRNA transport to the cytoplasm, and translation. About 20 major hnRNPs from A1 to U are known. Among them, hnRNP A, D, E, I, and K are known to shuttle between the nucleus and the cytoplasm. hnRNP E2 has been seen to stabilize α-globin mRNA and to enhance polioviral mRNA translation. hnRNP K modulates transcription and translation of some mRNAs. hnRNP I and its homologue hnRNP L have been suggested to enhance translation of some IRES-dependent mRNAs. In order to better understand the molecular mechanisms of the biological functions of hnRNPs, we investigated protein-protein interactions of six hnRNPs (hnRNP A1, C1, E2, I, K, and L) using the yeast two-hybrid system and in vitro co-precipitation assays. All of the hnRNPs tested exerted homomeric interactions, and hnRNP E2, I, K, and L interacted with each other. In the case of hnRNP E2 and hnRNP K, the N-terminal half of the proteins containing two KH (K homologous) domains were required for protein-protein interaction, and the second quarter of hnRNP I and hnRNP L containing RRM2 (RNA recognition motif 2) was essential for protein-protein interaction. hnRNP A1 and C1 did not form complexes with other hnRNPs in our assay systems. This suggests that the hnRNPs could fall into two groups: one group, including hnRNP A1 and C1, involved in hnRNP core complex formation and another group, including hnRNP E2, I, K, and L, involved in a variety of RNA-related biological processes. Different combinations of the proteins of the second group may facilitate different biological processes in conjunction with other factors.
Author Keywords: shuttling hnRNPs; protein-protein interaction; yeast two-hybrid; in vitro co-precipitation; RNA-related processes
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