Identifying small molecule inhibitors of eukaryotic translation initiation.
In eukaryotes, translation initiation is rate-limiting with much regulation exerted at the ribosome recruitment and ternary complex (eIF2.GTP.Met-tRNA(i)(Met)) formation steps. Although small molecule inhibitors have been extremely useful for chemically dissecting translation, there is a dearth of compounds available to study the initiation phase in vitro and in vivo. In this chapter, we describe reverse and forward chemical genetic screens developed to identify new inhibitors of translation. The ability to manipulate cell extracts biochemically, and to compare the activity of small molecules on translation of mRNA templates that differ in their factor requirements for ribosome recruitment, facilitates identification of the relevant target.
Cencic
Regina
R
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Robert
Francis
F
Pelletier
Jerry
J
D000465:Algorithms; D000818:Animals; D002499:Centrifugation, Density Gradient; D019411:Clinical Laboratory Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D008954:Models, Biological; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational / Q000187:drug effects*; D010448:Peptide Initiation Factors / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000302:isolation & purification*; D039381:RNA Cap-Binding Proteins / Q000378:metabolism / Q000502:physiology; D016601:RNA-Binding Proteins / Q000032:analysis; D011994:Recombinant Proteins / Q000138:chemical synthesis; D012107:Research Design; D012270:Ribosomes / Q000378:metabolism; D054852:Small Molecule Libraries / Q000032:analysis*
D010448:Peptide Initiation Factors; D011500:Protein Synthesis Inhibitors; D039381:RNA Cap-Binding Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D054852:Small Molecule Libraries