Primary cilia are key regulators of embryo development and tissue homeostasis. However, their mechanisms and functions, particularly in the context of human cells, are still unclear. Here, we analyzed the consequences of primary cilia modulation for human pluripotent stem cells (hPSCs) proliferation and differentiation. We report that neither activation of the cilia-associated Hedgehog signaling pathway nor ablation of primary cilia by CRISPR gene editing to knockout Tau Tubulin Kinase 2 (TTBK2), a crucial ciliogenesis regulator, affects the self-renewal of hPSCs. Further, we show that TTBK1, a related kinase without previous links to ciliogenesis, is upregulated during hPSCs-derived neural rosette differentiation. Importantly, we demonstrate that while TTBK1 fails to localize to the mother centriole, it regulates primary cilia formation in the differentiated, but not the undifferentiated hPSCs. Finally, we show that TTBK1/2 and primary cilia are implicated in the regulation of the size of hPSCs-derived neural rosettes.

• MeSH
• centrioly metabolismus   MeSH
• cilie metabolismus   MeSH
• lidé MeSH
• pluripotentní kmenové buňky * metabolismus   MeSH
• protein-serin-threoninkinasy genetika   metabolismus   MeSH
• proteiny hedgehog * genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• protein-serin-threoninkinasy MeSH
• proteiny hedgehog * MeSH
• tau-tubulin kinase MeSH Prohlížeč

Primary cilia play critical roles in development and disease. Their assembly and disassembly are tightly coupled to cell cycle progression. Here, we present data identifying KIF14 as a regulator of cilia formation and Hedgehog (HH) signaling. We show that RNAi depletion of KIF14 specifically leads to defects in ciliogenesis and basal body (BB) biogenesis, as its absence hampers the efficiency of primary cilium formation and the dynamics of primary cilium elongation, and disrupts the localization of the distal appendage proteins SCLT1 and FBF1 and components of the IFT-B complex. We identify deregulated Aurora A activity as a mechanism contributing to the primary cilium and BB formation defects seen after KIF14 depletion. In addition, we show that primary cilia in KIF14-depleted cells are defective in response to HH pathway activation, independently of the effects of Aurora A. In sum, our data point to KIF14 as a critical node connecting cell cycle machinery, effective ciliogenesis, and HH signaling.

• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• Aurora kinasa A antagonisté a inhibitory   genetika   metabolismus   MeSH
• bazální tělíska metabolismus   MeSH
• buněčný cyklus genetika   MeSH
• chromatografie kapalinová MeSH
• cilie genetika   metabolismus   patologie   MeSH
• HEK293 buňky MeSH
• interfáze fyziologie   MeSH
• intracelulární signální peptidy a proteiny genetika   metabolismus   MeSH
• kineziny genetika   metabolismus   MeSH
• lidé MeSH
• mitóza genetika   MeSH
• onkogenní proteiny genetika   metabolismus   MeSH
• protein-serin-threoninkinasy genetika   metabolismus   MeSH
• proteiny hedgehog metabolismus   MeSH
• RNA interference MeSH
• signální transdukce genetika   MeSH
• sodíkové kanály metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• AURKA protein, human MeSH Prohlížeč
• Aurora kinasa A MeSH
• citron-kinase MeSH Prohlížeč
• FBF1 protein, human MeSH Prohlížeč
• intracelulární signální peptidy a proteiny MeSH
• KIF14 protein, human MeSH Prohlížeč
• kineziny MeSH
• onkogenní proteiny MeSH
• protein-serin-threoninkinasy MeSH
• proteiny hedgehog MeSH
• SCLT1 protein, human MeSH Prohlížeč
• sodíkové kanály MeSH