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First published online 16 October 2007
doi: 10.1242/jcs.007955

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Tousled-like kinase in a microbial eukaryote regulates spindle assembly and S-phase progression by interacting with Aurora kinase and chromatin assembly factors

Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158-2280, USA

^* Author for correspondence (e-mail: ccwang{at}cgl.ucsf.edu)

Accepted 27 August 2007

The Tousled-like kinases are an evolutionarily conserved family of proteins implicated in DNA repair, DNA replication and mitosis in metazoans and plants. Their absence from the yeasts and other eukaryotic `microbes' suggests a specific role for them in the development of multicellular organisms. In this study, two closely related Tousled-like kinase homologs, TLK1 and TLK2, were identified in Trypanosoma brucei, a unicellular protozoan parasite. Only TLK1 plays an essential role in cell growth, and a deficiency in TLK1 led to an enrichment of S-phase cells, defective spindle formation and aberrant chromosome segregation. Although both TLK proteins localize to the nucleus, only TLK1 also concentrates in the spindle poles during mitosis. Both TLK proteins are phosphorylated by the Aurora kinase (AUK1), and both can autophosphorylate and phosphorylate histone H3 and the chromatin assembly factors Asf1A and Asf1B in vitro, but only TLK1 is autophosphorylated and capable of oligomerizing and interacting with AUK1, Asf1A and Asf1B in vivo. These discrepancies between the two TLK proteins can be attributed to minor differences between their N- and C-terminal sequences. In summary, TLK1 cooperates with Aurora kinase to regulate spindle assembly and chromosome segregation, and it performs a role in DNA replication probably by regulating histone modification in trypanosomes.

Key words: Anti-silencing function protein, Tousled-like kinase, Trypanosoma brucei, Mitosis, Spindle

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