Methods for identifying compounds that specifically target translation.
This chapter presents methods and protocols suitable for the identification and characterization of inhibitors of the prokaryotic and/or eukaryotic translational apparatus as a whole or targeting specific, underexploited targets of the bacterial protein synthetic machinery such as translation initiation and aminoacylation. Some of the methods described have been used successfully for the high-throughput screening of libraries of natural or synthetic compounds and make use of model "universal" mRNAs that can be translated with similar efficiency by cellfree extracts of bacterial, yeast, and HeLa cells. Other methods presented here are suitable for secondary screening tests aimed at identifying a specific target of an antibiotic within the translational pathway of prokaryotic cells.
Brandi
Letizia
L
Biotechnomics, Insubrias BioPark, Gerenzano, Italy.
Fabbretti
Attilio
A
Milon
Pohl
P
Carotti
Marcello
M
Pon
Cynthia L
CL
Gualerzi
Claudio O
CO
D000818:Animals; D001419:Bacteria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D002474:Cell-Free System / Q000378:metabolism; D002478:Cells, Cultured; D019411:Clinical Laboratory Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D049410:Luciferases, Renilla / Q000235:genetics / Q000378:metabolism; D019384:Nucleic Acid Synthesis Inhibitors / Q000032:analysis / Q000302:isolation & purification*; D039881:Prokaryotic Initiation Factor-2 / Q000037:antagonists & inhibitors / Q000502:physiology; D014176:Protein Biosynthesis / Q000187:drug effects*; D039381:RNA Cap-Binding Proteins / Q000502:physiology; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D046249:Transfer RNA Aminoacylation / Q000187:drug effects; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D019384:Nucleic Acid Synthesis Inhibitors; D039881:Prokaryotic Initiation Factor-2; D039381:RNA Cap-Binding Proteins; D012333:RNA, Messenger; D049410:Luciferases, Renilla