Ciliogenesis Dotaz Zobrazit nápovědu

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 insight into the substrates-TTBK2 kinase relationship and suggests that phosphorylation of substrates on multiple sites by TTBK2 is probably involved in the control of ciliogenesis in human cells.

MeSH
aminokyselinové motivy MeSH
cilie metabolismus MeSH
fosforylace MeSH
fosfoserin metabolismus MeSH
fosfothreonin metabolismus MeSH
HEK293 buňky MeSH
kaseinkinasa I metabolismus MeSH
lidé MeSH
multiproteinové komplexy metabolismus MeSH
organogeneze * MeSH
protein-serin-threoninkinasy chemie metabolismus MeSH
substrátová specifita MeSH
Check Tag
lidé MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Článek

KIF14 controls ciliogenesis via regulation of Aurora A and is important for Hedgehog signaling

We show that RNAi depletion of KIF14 specifically leads to defects in ciliogenesis and basal body (BB ...

The Journal of cell biology. 2020 ; 219 (6) : . [pub] 20200601

J Cell Biol
ISSN 1540-8140
Medvik
Zdroj

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.

Článek

Exon junction complex dependent mRNA localization is linked to centrosome organization during ciliogenesis

and immortalized human retinal pigment epithelial (RPE1) cells, centrioles form a basal body for ciliogenesis ...

Nature communications. 2021 ; 12 (1) : 1351. [pub] 20210301

Nat Commun
ISSN 2041-1723
Medvik
Zdroj

Exon junction complexes (EJCs) mark untranslated spliced mRNAs and are crucial for the mRNA lifecycle. An imbalance in EJC dosage alters mouse neural stem cell (mNSC) division and is linked to human neurodevelopmental disorders. In quiescent mNSC and immortalized human retinal pigment epithelial (RPE1) cells, centrioles form a basal body for ciliogenesis. Here, we report that EJCs accumulate at basal bodies of mNSC or RPE1 cells and decline when these cells differentiate or resume growth. A high-throughput smFISH screen identifies two transcripts accumulating at centrosomes in quiescent cells, NIN and BICD2. In contrast to BICD2, the localization of NIN transcripts is EJC-dependent. NIN mRNA encodes a core component of centrosomes required for microtubule nucleation and anchoring. We find that EJC down-regulation impairs both pericentriolar material organization and ciliogenesis. An EJC-dependent mRNA trafficking towards centrosome and basal bodies might contribute to proper mNSC division and brain development.

MeSH
autoantigeny metabolismus MeSH
buněčný cyklus MeSH
centrozom metabolismus MeSH
cilie metabolismus MeSH
cytoskeletální proteiny metabolismus MeSH
DEAD-box RNA-helikasy metabolismus MeSH
eukaryotický iniciační faktor 4A metabolismus MeSH
exony genetika MeSH
jaderné proteiny metabolismus MeSH
lidé MeSH
messenger RNA metabolismus MeSH
mikrotubuly metabolismus MeSH
myši MeSH
nervové kmenové buňky metabolismus MeSH
proliferace buněk MeSH
proteiny asociované s mikrotubuly metabolismus MeSH
proteiny buněčného cyklu metabolismus MeSH
proteiny vázající RNA metabolismus MeSH
proteosyntéza MeSH
transport RNA * MeSH
zvířata MeSH
Check Tag
lidé MeSH
myši MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Článek

Tau tubulin kinase 1 and 2 regulate ciliogenesis and human pluripotent stem cells-derived neural rosettes

ablation of primary cilia by CRISPR gene editing to knockout Tau Tubulin Kinase 2 (TTBK2), a crucial ciliogenesis ...

Scientific reports. 2023 ; 13 (1) : 12884. [pub] 20230809

Sci Rep
ISSN 2045-2322
Medvik
Zdroj

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.