title: Development of a chemical genetic approach for human aurora B kinase identifies novel substrates of the chromosomal passenger complex.
abstract: To understand how the chromosomal passenger complex ensures chromosomal stability, it is crucial to identify its substrates and to find ways to specifically inhibit the enzymatic core of the complex, Aurora B. We therefore developed a chemical genetic approach to selectively inhibit human Aurora B. By mutating the gatekeeper residue Leu-154 in the kinase active site, the ATP-binding pocket was enlarged, but kinase function was severely disrupted. A unique second site suppressor mutation was identified that rescued kinase activity in the Leu-154 mutant and allowed the accommodation of bulky N(6)-substituted adenine analogs. Using this analog-sensitive Aurora B kinase, we found that retention of the chromosomal passenger complex at the centromere depends on Aurora B kinase activity. Furthermore, analog-sensitive Aurora B was able to use bulky ATPγS analogs and could thiophosphorylate multiple proteins in cell extracts. Utilizing an unbiased approach for kinase substrate mapping, we identified several novel substrates of Aurora B, including the nucleosomal-binding protein HMGN2. We confirmed that HMGN2 is a bona fide Aurora B substrate in vivo and show that its dynamic association to chromatin is controlled by Aurora B.
authors.affiliation: Department of Medical Oncology, University Medical Center, 3584 CG Utrecht, The Netherlands.
authors.forename: Rutger C C
authors.initials: RC
authors.lastname: Hengeveld
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Hertz
authors.forename: Martijn J M
authors.initials: MJ
authors.lastname: Vromans
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.forename: Susanne M A
authors.initials: SM
authors.lastname: Lens
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D064107:Aurora Kinase B; D064127:Aurora Kinases; D020134:Catalytic Domain; D002457:Cell Extracts / Q000737:chemistry; D045744:Cell Line, Tumor; D002503:Centromere; D017124:Conserved Sequence; D011756:Diphosphates / Q000494:pharmacology; D024242:HMGN2 Protein / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D007930:Leucine / Q000235:genetics; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D002457:Cell Extracts; D011756:Diphosphates; D024242:HMGN2 Protein; D006657:Histones; D046912:Multiprotein Complexes; D010750:Phosphoproteins; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D007930:Leucine