Kinesin-13 regulates the quantity and quality of tubulin inside cilia

Vasudevan, Krishna Kumar; Jiang, Yu-Yang; Lechtreck, Karl‐Ferdinand; Kushida, Yasuharu; Alford, Lea M.; Sale, Winfield S.; Hennessey, Todd M.; Gaertig, Jacek

doi:10.1091/mbc.e14-09-1354

Cited by 31 publications

(34 citation statements)

References 82 publications

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“…S4). These results, therefore, indicated the presence of KATNAL2 at the cilium; to date the localization of katanin and MT-depolymerising kinesin-like proteins have been documented at the ciliary axoneme and basal body [14,31,32,[46][47][48] and thus KATNAL2 proteins can now be added as putative players in axonemal dynamics.…”

Section: Katnal2 Isoforms Interact With Themselves With Each Other Amentioning

confidence: 63%

A novel family of katanin-like 2 protein isoforms (KATNAL2), interacting with nucleotide-binding proteins Nubp1 and Nubp2, are key regulators of different MT-based processes in mammalian cells

Ververis

Christodoulou

Christoforou

et al. 2015

Cell. Mol. Life Sci.

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Katanins are microtubule (MT)-severing AAA proteins with high phylogenetic conservation throughout the eukaryotes. They have been functionally implicated in processes requiring MT remodeling, such as spindle assembly in mitosis and meiosis, assembly/disassembly of flagella and cilia and neuronal morphogenesis. Here, we uncover a novel family of katanin-like 2 proteins (KAT-NAL2) in mouse, consisting of five alternatively spliced isoforms encoded by the Katnal2 genomic locus. We further demonstrate that in vivo these isoforms are able to interact with themselves, with each other and moreover directly and independently with MRP/MinD-type P-loop NTPases Nubp1 and Nubp2, which are integral components of centrioles, negative regulators of ciliogenesis and implicated in centriole duplication in mammalian cells. We find KATNAL2 localized on interphase MTs, centrioles, mitotic spindle, midbody and the axoneme and basal body of sensory cilia in cultured murine cells. shRNAi of Katnal2 results in inefficient cytokinesis and severe phenotypes of enlarged cells and nuclei, increased numbers of centrioles and the manifestation of aberrant multipolar mitotic spindles, mitotic defects, chromosome bridges, multinuclearity, increased MT acetylation and an altered cell cycle pattern. Silencing or stable overexpression of KATNAL2 isoforms drastically reduces ciliogenesis. In conclusion, KATNAL2s are multitasking enzymes involved in the same cell type in critically important processes affecting cytokinesis, MT dynamics, and ciliogenesis and are also implicated in cell cycle progression.

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Section: Katnal2 Isoforms Interact With Themselves With Each Other Amentioning

confidence: 63%

A novel family of katanin-like 2 protein isoforms (KATNAL2), interacting with nucleotide-binding proteins Nubp1 and Nubp2, are key regulators of different MT-based processes in mammalian cells

Ververis

Christodoulou

Christoforou

et al. 2015

Cell. Mol. Life Sci.

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“…The balance point model of flagellar length control requires length dependence of either assembly and disassembly rates to achieve overall length control (Mohapatra et al, 2016). In Chlamydomonas and Tetrahymena length control is achieved through length-dependent IFT assembly with length-independent disassembly (Engel et al, 2009; Vasudevan et al, 2015). As we were unable to confirm length-dependent IFT-mediated assembly for the four Giardia flagellar pairs, we also investigated the potential contribution of length-dependent disassembly to the maintenance of four different equilibrium lengths.…”

Section: Resultsmentioning

confidence: 99%

“…Kinesin-13 contributions to flagellar assembly and disassembly have also been investigated in other microbial flagellates such as Leishmania, Tetrahymena, and Chlamydomonas (Blaineau et al, 2007; Chan & Ersfeld, 2010; Li et al, 2009; Vasudevan et al, 2015; Wang et al, 2013). Kinesin-13 has a similar role in mediating axoneme disassembly in Leishmania , where the knockdown of one (of the seven) kinesin-13 homologs promotes increased flagellar length, and overexpression of the same kinesin-13 homolog results decreased flagellar length (Blaineau et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

Length-dependent disassembly maintains four different flagellar lengths in Giardia

McInally

Kondev

Dawson

2019

Preprint

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1How flagellar length regulation is achieved in multiciliated eukaryotic cells with flagella of 2 different equilibrium lengths is unknown. The protist Giardia lamblia is an ideal model to 3 evaluate length regulation as it has flagella of four different lengths. Giardia axonemes have 4 both non-membrane-bound and membrane-bound regions, but lack transition zones. Here we 5 quantified the contributions of intraflagellar transport (IFT)-mediated assembly and kinesin-13-6 mediated disassembly to length control. IFT particles assemble and inject at Giardia's flagellar 7 pore complexes, which act as diffusion barriers functionally analogous to the transition zone to 8 compartmentalize the membrane-bound regions of flagella. IFT-mediated assembly is length-9 independent as train size, speed, and injection frequencies are similar between flagella of 10 different lengths. In Giardia, kinesin-13 mediates a length-dependent disassembly mechanism 11 of length regulation to balance length-independent IFT-mediated assembly, resulting in 12 different lengths. We anticipate that similar control mechanisms are widespread in multiciliated 13 cells where cytoplasmic precursor pools are not limiting. 14 15 16 17

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“…SPP medium [111] with antibiotics, SPPA [112] was used to grow Tetrahymena with exception of strains with severe cilia defects, which were maintained in MEPP medium [89] with 2 µg/ml dextrose (MEPPD) [25]. CdCl 2 (2.5 µg/ml) was added to induce MTT1-driven overproduction of proteins [73].…”

Section: Methodsmentioning

confidence: 99%

LF4/MOK and a CDK-related kinase regulate the number and length of cilia inTetrahymena

Jiang

Maier

Baumeister

et al. 2019

Preprint

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22The length of cilia is controlled by a poorly understood mechanism that involves 23 members of the conserved RCK kinase group, and among them, the LF4/MOK 24 kinases. In Tetrahymena, a loss of an LF4/MOK ortholog, LF4A, lengthened the 25 locomotory cilia, but also reduced their total number per cell. Without LF4A, cilia 26 assembled faster and showed signs of increased intraflagellar transport (IFT). 27Consistently, overproduced LF4A shortened cilia and downregulated the IFT. GFP-28 tagged LF4A, expressed in the native locus and imaged by total internal reflection 29 microscopy, was enriched at the basal bodies and distributed along the shafts of 30 cilia. Within cilia, most LF4A-GFP particles were immobile and a few either diffused or 31 moved by IFT. A forward genetic screen identified a CDK-related kinase, CDKR1, 32 whose loss-of-function suppressed the shortening of cilia caused by overexpression 33 of LF4A, by reducing its kinase activity. A loss of CDKR1 alone lengthened both the 34 locomotory and oral cilia. CDKR1 resembles other known ciliary CDK-related kinases: 35 LF2 of Chlamydomonas, mammalian CCRK and DYF-18 of C. elegans, in lacking the 36 cyclin-binding motif and acting upstream of RCKs. We propose that the total 37 LF4/MOK activity per cilium is dependent on both its activation by an upstream CDK-38 related kinase and cilium length. Previous studies showed that the rate of assembly 39 is high in growing cilia and decreases as cilia elongate to achieve the steady-state 40 length. We propose that in a longer cilium, the IFT components, which travel from 41 the base to the tip, are subjected to a higher dose of inhibition by the uniformly 42 3 distributed LF4/MOK. Thus, in a feedback loop, LF4/MOK may translate cilium length 43 into proportional inhibition of IFT, to balance the rates of assembly and disassembly 44 at steady-state. 45 46Author summary 47 Cilia are conserved organelles that generate motility and mediate vital sensory 48 functions, including olfaction and vision. Cilia that are either too short or too long fail 49 to generate proper forces or responses to extracellular signals. Several cilia-based 50 diseases (ciliopathies) are associated with defects in cilia length. Here we use the 51 multiciliated model protist Tetrahymena, to study a conserved protein kinase whose 52 activity shortens cilia, LF4/MOK. We find that cells lacking an LF4/MOK kinase of 53Tetrahymena, LF4A, have excessively long, but also fewer cilia. We show that LF4A 54 decreases the intraflagellar transport, a motility that shuttles ciliary precursors from 55 the cilium base to the tip. Live imaging revealed that LF4A is distributed along cilium 56 length and remains mostly immobile, likely due to its anchoring to ciliary 57 microtubules. We proposed that in longer cilia, the intraflagellar transport machinery 58 is exposed to a higher dose of inhibition by LF4A, which could decrease the rate of 59 cilium assembly, to balance the rate of cilium disassembly in mature cilia that 60 maintain stable length. 61 62 63T...

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Kinesin-13 regulates the quantity and quality of tubulin inside cilia

Cited by 31 publications

References 82 publications

A novel family of katanin-like 2 protein isoforms (KATNAL2), interacting with nucleotide-binding proteins Nubp1 and Nubp2, are key regulators of different MT-based processes in mammalian cells

A novel family of katanin-like 2 protein isoforms (KATNAL2), interacting with nucleotide-binding proteins Nubp1 and Nubp2, are key regulators of different MT-based processes in mammalian cells

Length-dependent disassembly maintains four different flagellar lengths in Giardia

LF4/MOK and a CDK-related kinase regulate the number and length of cilia inTetrahymena

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