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Identification of cellular proteins enhancing activities of internal ribosomal entry sites by competition with oligodeoxynucleotides

By Kobong Choi, Jong Heon Kim, Xiaoyu Li, Ki Young Paek, Sang Hoon Ha, Sung Ho Ryu, Eckard Wimmer and Sung Key Jang

Abstract

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

Topics: Articles
Publisher: Oxford University Press
Year: 2004
DOI identifier: 10.1093/nar/gkh300
OAI identifier: oai:pubmedcentral.nih.gov:390288
Provided by: PubMed Central
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