The Ipl1/Aurora kinase family: methods of inhibition and functional analysis in mammalian cells.
The Ipl1/Aurora family of protein kinases are required for accurate chromosome segregation. Because members of this family are often overexpressed in human tumors, they have recently received much attention, both from the academic community and the pharmaceutical industry. Indeed, two small molecule Aurora kinase inhibitors have recently been described. In this chapter, we describe several methods for investigating the function of the Aurora kinases, focusing on Aurora B. We describe the use of the small-molecule inhibitor ZM447439, RNA interference, and overexpression of a catalytic mutant. All of these methods have proved useful in studying Aurora B as well as validating it as a potential anticancer drug target. However, while all three methods are useful for probing the function of Aurora B, each has inherent advantages and disadvantages. Furthermore, because the mechanism underlying the inhibition is different in each case, caution must be taken when interpreting the data.
Ditchfield
Claire
C
School of Biological Sciences, University of Manchester, Manchester, UK.
Keen
Nicholas
N
Taylor
Stephen S
SS
D064107:Aurora Kinase B; D064127:Aurora Kinases; D001483:Base Sequence; D001549:Benzamides / Q000494:pharmacology; D020134:Catalytic Domain / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000032:analysis* / Q000037:antagonists & inhibitors* / Q000235:genetics; D011799:Quinazolines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D014162:Transfection
C474722:4-(4-(N-benzoylamino)anilino)-6-methoxy-7-(3-(1-morpholino)propoxy)quinazoline; D001549:Benzamides; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases