tailieunhanh - Báo cáo khoa học: Identification of Ewing’s sarcoma protein as a G-quadruplex DNA- and RNA-binding protein

The Ewing’s sarcoma (EWS) oncogene contains an N-terminal transcrip-tion activation domain and a C-terminal RNA-binding domain. Although the EWS activation domain is a potent transactivation domain that is required for the oncogenic activity of several EWS fusion proteins, the nor-mal role of intact EWS is poorly characterized because little is known about its nucleic acid recognition specificity. | 1FEBS Journal Identification of Ewing s sarcoma protein as a G-quadruplex DNA- and RNA-binding protein Kentaro Takahama1 Katsuhito Kino2 Shigeki Arai3 Riki Kurokawa3 and Takanori Oyoshi1 1 Department of Chemistry Faculty of Science Shizuoka University Japan 2 Kagawa Schoolof PharmaceuticalSciences Tokushima Bunri University Kagawa Japan 3 Division of Gene Structure and Function Saitama MedicalUniversity Research Center for Genomic Medicine Japan Keywords Ewing s sarcoma G-quadruplex DNA G-quadruplex RNA RGG motif RNA-binding protein Correspondence T. Oyoshi Department of Chemistry Faculty of Science Graduate School of Science Shizuoka University 836 Oya Suruga Shizuoka 422-8529 Japan Fax 81 54 237 3384 Tel 81 54 238 4760 E-mail stohyos@ These authors contributed equally to this work Received 27 August 2010 revised 23 December 2010 accepted 13 January 2011 doi The Ewing s sarcoma EWS oncogene contains an N-terminal transcription activation domain and a C-terminal RNA-binding domain. Although the EWS activation domain is a potent transactivation domain that is required for the oncogenic activity of several EWS fusion proteins the normal role of intact EWS is poorly characterized because little is known about its nucleic acid recognition specificity. Here we show that the Arg-Gly-Gly RGG domain of the C-terminal in EWS binds to the G-rich single-stranded DNA and RNA fold in the G-quadruplex structure. Furthermore inhibition of DNA polymerase on a template containing a human telomere sequence in the presence of RGG occurs in an RGG concentration-dependent manner by the formation of a stabilized G-quad-ruplex DNA-RGG complex. In addition mutated RGG containing Lys residues replacing Arg residues at specific Arg-Gly-Gly sites and RGG containing Arg methylated by protein arginine N-methyltransferase 3 decrease the binding ability of EWS to G-quadruplex DNA and RNA. These findings suggest that the RGG of EWS binds to .