TPX2 Protein of Arabidopsis Activates Aurora Kinase 1, But Not Aurora Kinase 3 In Vitro
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Aurora kinases are involved in various mitotic events, including chromosome segregation and bipolar mitotic spindle assembly. In animals, Aurora A is activated and protected by microtubule-associated protein TPX2. Such role in plants is not known. Here, we have assessed the ability of TPX2 of Arabidopsis to regulate Aurora family members in vitro. AtTPX2 acts as substrate as well as activator of AtAurora1, but not AtAurora3. Truncated version of AtTPX2 lacking the Aurora binding domain is unable to activate the kinases; however, it is still phosphorylated. AtTPX2-induced activation of AtAurora1 results in a dramatically increased phosphorylation level of downstream targets, particularly histone H3. The differences in the activation mechanism of AtAurora1 and 3 point to a specific regulation of both kinases, which may play an important role in cell cycle regulation and signaling cascade transduction in plants.
KeywordsAurora kinase Targeting protein for Xklp2 In vitro kinase assay Kinase activation
We thank Oda Weiss for the excellent technical support. This research was supported by grants from the Czech Science Foundation (14-28443S) and the National Program of Sustainability I (LO1204); Internal Grant Agency of Palacky University, Olomouc (IGA_PrF_2014001, PrF-2013-003) and Interdoc (OPVK-CZ.1.07/2.4.00/17.0008) for ET and HJ. DD and AH are supported by the DFG (SFB 648). Grant from the Czech Science Foundation P501-12-2333 for PB. We thank Dr. H. P. Mock for helpful discussion about protein purification.
- Kurihara D, Sachihiro M, Uchiyama S, Fukui K (2008) Live cell imaging reveals plant aurora kinase has dual roles during mitosis. Plant Cell Physiol 49:1256–1261Google Scholar
- Neumayer G, Helfricht A, Shim SY, Le HT, Lundin C, Belzil C, Chansard M, Yu Y, Lees-Miller SP, Gruss OJ, can Attikum H, Helleday T, Nguyen MD (2012) Targeting protein for xenopus kinesin-like protein 2 (TPX2) regulates gamma histone 2AX (gamma-H2AX) levels upon ionizing radiation. J Biol Chem 287:42206–42222PubMedPubMedCentralCrossRefGoogle Scholar
- Petrovská B, Jeřábková H, Chamrád I, Vrána J, Lenobel R, Uřinovská J, Šebela M, Doležel J (2014) Proteomic analyses of barley cell nuclei purified by flow sorting. Cytogenet. Genome Res 143:78–86Google Scholar
- Petrovská B, Jeřábková H, Kohoutová L, Cenklová V, Pochylová Ž, Gelová Z, Kočárová G, Váchová L, Kurejová M, Tomaštíková E, Binarová P (2013) Overexpressed TPX2 causes ectopic formation of microtubular arrays in the nuclei of acentrosomal plant cells. J Exp Bot 64:4575–4587PubMedPubMedCentralCrossRefGoogle Scholar