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- W2027982315 abstract "Eukaryotic initiation factor eIF4A is a prototype protein of the DEAD box family of RNA helicases, and is part of the translation initiation complex eIF4F 1.eIF4A binds to the 5′ cap of mRNA and unwinds structures in the 5′-untranslated regions of mRNAs in ATP dependent manner. Although eIF4A has been studied extensively by classical bulk biochemical methods, a direct, unambiguous measurement of its helicase activity and its processivity has not been reported. Here, we use single molecule fluorescence assays to visualize its binding to RNA and melting secondary structures in RNA. Specifically, FRET efficiency dynamics is used to explore the binding location of eIF4A and its unwinding function. Our single molecule studies show that eIF4A has higher binding affinity towards the duplex site. We seek to elucidate any elementary steps and kinetic mechanisms involved with eIF4A unwinding of RNA. Furthermore, we selectively target eIF4A activity with small-molecule inhibitors acting in opposite manners2. The dynamics of stimulation and inhibition of eIF4A activity by pateamine and hippuristanol are measured at the single molecule level. The results will provide insight into the eIF4A's helicase activity and will distinguish between passive versus active unwinding mechanism. Our long-term goal in this project is to decipher the role of the initiation complex eIF4F in ribosomal recruitment, and develop methods to control this process. Then, we will probe eIF4A activity in the presence of other initiation (co)factors. 1. Rogers, G. W., Jr., Komar, A. A. & Merrick, W. C. eIF4A: The godfather of the DEAD box helicases. Prog Nucleic Acid Res Mol Biol 72, 307-331 (2002). 2. Bordeleau, M. E. et al. Stimulation of mammalian translation initiation factor eIF4A activity by a small molecule inhibitor of eukaryotic translation. Proc Natl Acad Sci. 102, 10460-5 (2005)." @default.
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- W2027982315 date "2009-02-01" @default.
- W2027982315 modified "2023-09-26" @default.
- W2027982315 title "Deciphering the Mechanism of RNA helicase eIF4A in Translation Initiation" @default.
- W2027982315 doi "https://doi.org/10.1016/j.bpj.2008.12.2121" @default.
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