Phosphorylation of multiple proteins involved in ciliogenesis by Tau Tubulin kinase 2

Molecular Biology of the Cell
Ondrej BernatikLukas Cajanek

Abstract

Primary cilia are organelles necessary for proper implementation of developmental and homeostasis processes. To initiate their assembly, coordinated actions of multiple proteins are needed. Tau tubulin kinase 2 (TTBK2) is a key player in the cilium assembly pathway, controlling the final step of cilia initiation. The function of TTBK2 in ciliogenesis is critically dependent on its kinase activity; however, the precise mechanism of TTBK2 action has so far not been fully understood due to the very limited information about its relevant substrates. In this study, we demonstrate that CEP83, CEP89, CCDC92, Rabin8, and DVL3 are substrates of TTBK2 kinase activity. Further, we characterize a set of phosphosites of those substrates and CEP164 induced by TTBK2 in vitro and in vivo. Intriguingly, we further show that identified TTBK2 phosphosites and consensus sequence delineated from those are distinct from motifs previously assigned to TTBK2. Finally, we show that TTBK2 is also required for efficient phosphorylation of many S/T sites in CEP164 and provide evidence that TTBK2-induced phosphorylations of CEP164 modulate its function, which in turn seems relevant for the process of cilia formation. In summary, our work provides important ...Continue Reading

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Citations

Mar 16, 2021·Frontiers in Cell and Developmental Biology·Ondrej BernatikLukas Cajanek
Apr 30, 2021·Frontiers in Cell and Developmental Biology·Elena A MayDavid U Mick
May 7, 2021·The Journal of Cell Biology·Tiphaine DouanneGillian M Griffiths

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Methods Mentioned

BETA
protein
immunoprecipitation
protein folding
transfection
Electrophoresis
PCR

Software Mentioned

PONDR VSL2
Mascot
MoMo simple
MoMo
MeMe
Predictor of ( PONDR
Graphpad Prism
DEPP
Photoshop
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