Phosphosite <scp>T674A</scp> mutation in kinesin family member <scp>3A</scp> fails to reproduce tissue and ciliary defects characteristic of <scp>CILK1</scp> loss of function

Gailey, Casey D.; Wang, Eric J.; Jin, Li; Ahmadi, Sean; Brautigan, David L.; Li, Xudong; Xu, Wenhao; Scott, Michael M.; Fu, Zheng

doi:10.1002/dvdy.252

Cited by 15 publications

(9 citation statements)

References 35 publications

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“…A key phosphorylation target of RCK is heterotrimeric kinesin-2: in mammals KIF3A [ 58 , 79 ] and in Chlamydomonas FLA8, the homolog of KIF3B [ 177 ]. In mice, KIF3A has been shown to be not the only target of CILK1, since mutants expressing a non-phosphorylatable KIF3A variant have a distinct phenotype from CILK1-knockouts [ 178 ]. The phenotypes of the RCK-knockouts in different organisms–(tip) accumulation of IFT components– resemble those of dynein-2 and IFT-A knockouts [ 11 , 179 , 180 , 181 , 182 ].…”

Section: Ift Regulation From the Perspective Of Regulatory Proteinsmentioning

confidence: 99%

Mechanisms of Regulation in Intraflagellar Transport

Mul

Mitra

2022

Cells

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Cilia are eukaryotic organelles essential for movement, signaling or sensing. Primary cilia act as antennae to sense a cell’s environment and are involved in a wide range of signaling pathways essential for development. Motile cilia drive cell locomotion or liquid flow around the cell. Proper functioning of both types of cilia requires a highly orchestrated bi-directional transport system, intraflagellar transport (IFT), which is driven by motor proteins, kinesin-2 and IFT dynein. In this review, we explore how IFT is regulated in cilia, focusing from three different perspectives on the issue. First, we reflect on how the motor track, the microtubule-based axoneme, affects IFT. Second, we focus on the motor proteins, considering the role motor action, cooperation and motor-train interaction plays in the regulation of IFT. Third, we discuss the role of kinases in the regulation of the motor proteins. Our goal is to provide mechanistic insights in IFT regulation in cilia and to suggest directions of future research.

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Section: Ift Regulation From the Perspective Of Regulatory Proteinsmentioning

confidence: 99%

Mechanisms of Regulation in Intraflagellar Transport

Mul

Mitra

2022

Cells

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“…Mutations in the ICK gene in humans cause ciliopathies [34][35][36], and knockout (KO) and mutant Ick mice manifest ciliopathy phenotypes [32,37,38]. Previous studies proposed that the phosphorylation of KIF3A, a kinesin-II subunit, by ICK is crucial for the regulation of ciliary protein trafficking [32,39]; however, a recent study suggested that KIF3A phosphorylation is dispensable for ciliary function [40].…”

Section: Introductionmentioning

confidence: 99%

CCRK/CDK20 regulates ciliary retrograde protein trafficking via interacting with BROMI/TBC1D32

et al. 2021

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CCRK/CDK20 was reported to interact with BROMI/TBC1D32 and regulate ciliary Hedgehog signaling. In various organisms, mutations in the orthologs of CCRK and those of the kinase ICK/CILK1, which is phosphorylated by CCRK, are known to result in cilia elongation. Furthermore, we recently showed that ICK regulates retrograde ciliary protein trafficking and/or the turnaround event at the ciliary tips, and that its mutations result in the elimination of intraflagellar transport (IFT) proteins that have overaccumulated at the bulged ciliary tips as extracellular vesicles, in addition to cilia elongation. However, how these proteins cooperate to regulate ciliary protein trafficking has remained unclear. We here show that the phenotypes of CCRK-knockout (KO) cells closely resemble those of ICK-KO cells; namely, the overaccumulation of IFT proteins at the bulged ciliary tips, which appear to be eliminated as extracellular vesicles, and the enrichment of GPR161 and Smoothened on the ciliary membrane. The abnormal phenotypes of CCRK-KO cells were rescued by the exogenous expression of wild-type CCRK but not its kinase-dead mutant or a mutant defective in BROMI binding. These results together indicate that CCRK regulates the turnaround process at the ciliary tips in concert with BROMI and probably via activating ICK.

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“…Both KIF3A and GSK3β have been linked to intraflagellar transport and cilia length control [ 32 , 33 ]. However, our prior study showed that deficiency in CILK1 phosphorylation of KIF3A alone is insufficient to account for the cilia phenotype caused by CILK1 inactivation [ 34 ]. Therefore, it remains a mystery as to which CILK1 substrate(s) mediate the effect of Alvocidib in the regulation of intraflagellar transport and cilia length.…”

Section: Discussionmentioning

confidence: 99%

Modulation of Primary Cilia by Alvocidib Inhibition of CILK1

Wang

Turner

Brautigan

et al. 2022

IJMS

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The primary cilium provides cell sensory and signaling functions. Cilia structure and function are regulated by ciliogenesis-associated kinase 1 (CILK1). Ciliopathies caused by CILK1 mutations show longer cilia and abnormal Hedgehog signaling. Our study aimed to identify small molecular inhibitors of CILK1 that would enable pharmacological modulation of primary cilia. A previous screen of a chemical library for interactions with protein kinases revealed that Alvocidib has a picomolar binding affinity for CILK1. In this study, we show that Alvocidib potently inhibits CILK1 (IC50 = 20 nM), exhibits selectivity for inhibition of CILK1 over cyclin-dependent kinases 2/4/6 at low nanomolar concentrations, and induces CILK1-dependent cilia elongation. Our results support the use of Alvocidib to potently and selectively inhibit CILK1 to modulate primary cilia.

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Phosphosite T674A mutation in kinesin family member 3A fails to reproduce tissue and ciliary defects characteristic of CILK1 loss of function

Cited by 15 publications

References 35 publications

Mechanisms of Regulation in Intraflagellar Transport

Mechanisms of Regulation in Intraflagellar Transport

CCRK/CDK20 regulates ciliary retrograde protein trafficking via interacting with BROMI/TBC1D32

Modulation of Primary Cilia by Alvocidib Inhibition of CILK1

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