| Title | Identification of cellular proteins enhancing activities of internal ribosomal entry sites by competition with oligodeoxynucleotides. | ||
| Author | Choi, Kobong; Kim, Jong Heon; Li, Xiaoyu; Paek, Ki Young; Ha, Sang Hoon; Ryu, Sung Ho; Wimmer, Eckard; Jang, Sung Key | ||
| Journal | Nucleic Acids Res | Publication Year/Month | 2004 |
| PMID | 14981151 | PMCID | PMC390288 |
| Affiliation | 1.Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, San 31, Hyoja-Dong, Pohang, Kyungbuk 790-784, Korea. | ||
The translation of numerous eukaryotic mRNAs is mediated by internal ribosomal entry sites (IRESs). IRES-dependent translation requires both canonical translation initiation factors and IRES-specific trans-acting factors (ITAFs). Here we report a strategy to identify and characterize ITAFs required for IRES-dependent translation. This process involves steps for identifying oligodeoxynucleotides affecting IRES-dependent translation, purifying proteins interacting with the inhibitory DNA, identifying the specific proteins with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry, and confirming the roles of these proteins in IRES-dependent translation by depletion and repletion of proteins from an in vitro translation system. Using this strategy, we show that poly(rC)-binding proteins 1 and 2 enhance translation through polioviral and rhinoviral IRES elements.