Homogenous time resolved fluorescence assay to identify modulators of cap-dependent translation initiation.
Eukaryotic initiation factor (eIF) 4F plays a key role in recruiting 40S ribosomes and associated factors to mRNA templates during translation initiation. The function of this heterotrimeric complex is to deliver an RNA helicase to the 5' cap proximal region of mRNAs in preparation for ribosome binding. To study the interaction between subunits of this complex, as well as identify small molecules that could interfere with their association, we developed a time resolved fluorescence assay that allows monitoring of interactions between two subunits of eIF4F. We have performed a small molecule chemical screen of >73,000 compounds using this assay.
Cencic
Regina
R
Department of Biochemistry, McIntyre Medical Sciences Building, 3655 Promenade Sir William Osler, McGill University, Montreal, Quebec, Canada, H3G 1Y6.
Yan
Yifei
Y
Pelletier
Jerry
J
D004353:Drug Evaluation, Preclinical / Q000379:methods*; D039562:Eukaryotic Initiation Factor-4F / Q000037:antagonists & inhibitors*; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D014176:Protein Biosynthesis / Q000187:drug effects*; D021122:Protein Subunits / Q000037:antagonists & inhibitors; D012315:RNA Caps; D012270:Ribosomes / Q000378:metabolism
D039562:Eukaryotic Initiation Factor-4F; D021122:Protein Subunits; D012315:RNA Caps