The Aurora B kinase in Trypanosoma brucei undergoes post-translational modifications and is targeted to various subcellular locations through binding to TbCPC1.
The chromosomal passenger complex (CPC) in animals, consisting of Aurora B kinase and three evolutionarily conserved proteins, plays crucial roles in mitosis and cytokinesis. However, Trypanosoma brucei expresses an unusual CPC consisting of an Aurora-like kinase, TbAUK1, and two kinetoplastid-specific proteins, TbCPC1 and TbCPC2. Despite their essential functions, little is known about the regulation of TbAUK1 and the roles of TbCPC1 and TbCPC2. Here, we investigate the effect of post-translational modification on the activity and spatiotemporal control of TbAUK1, and demonstrate that phosphorylation of two conserved threonine residues in the activation loop of the kinase domain contributes to TbAUK1 activation and function. TbAUK1 is SUMOylated in vivo, and mutation of the SUMO-conjugation site compromises TbAUK1 function. Degradation of TbAUK1 requires two destruction boxes and is mediated by the anaphase-promoting complex/cyclosome (APC/C), whereas degradation of TbCPC1 and TbCPC2 is not dependent on the predicted destruction boxes and is APC/C-independent. Moreover, we determine the domains in CPC subunits that mediate the pairwise interactions, and show that disruption of the interaction impairs the localization of TbAUK1 and TbCPC2 but not TbCPC1. Our results demonstrate the requirement of post-translational modifications for TbAUK1 function and a crucial role of TbCPC1 in mediating TbAUK1 localization.
Department of Microbiology and Molecular Genetics, University of Texas Medical School at Houston, Houston, TX, 77030, USA.
Huiqing
H
Hu
Zhonglian
Z
Yu
Yi
Y
Liu
Tao
T
Wang
Ying
Y
Wei
Ziyin
Z
Li
D064107:Aurora Kinase B / Q000235:genetics* / Q000378:metabolism*; D020134:Catalytic Domain; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D009154:Mutation; D010766:Phosphorylation; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D055550:Protein Stability; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation; D013912:Threonine / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000166:cytology / Q000201:enzymology* / Q000235:genetics
D015800:Protozoan Proteins; D013912:Threonine; D064107:Aurora Kinase B