Centriole splitting caused by loss of the centrosomal linker protein C-NAP1 reduces centriolar satellite density and impedes centrosome amplification

Flanagan, Adrienne M.; Stavenschi, Elena; Basavaraju, Shivakumar; Gaboriau, David; Hoey, David A.; Morrison, Ciarán

doi:10.1091/mbc.e16-05-0325

Cited by 25 publications

(38 citation statements)

References 62 publications

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“…This observation suggests a role for centrin 2 in satellite regulation, consistent with the previously described localization of centrin to satellites (Löffler et al, 2013;Prosser et al, 2009). Alterations in centriolar satellite density can affect centrosome duplication (Flanagan et al, 2017;Kodani et al, 2015;Löffler et al, 2013;Prosser et al, 2009). Therefore, we quantitated centrosome amplification after ionizing (IR) and ultraviolet (UV) irradiation, visualizing centrioles with the proximal end marker CEP135 (Ohta et al, 2002) and centrin 3, which is representative of a distinct family of centrins ( Fig.…”

Section: Analysis Of the Ef-hand Domains Of Centrin 2 In Centriolar Lsupporting

confidence: 88%

Differential requirements for the EF-hand domains of human centrin 2 in primary ciliogenesis and nucleotide excision repair

Khouj

Prosser

Tada

et al. 2019

Journal of Cell Science

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Centrin 2 is a small conserved calcium-binding protein that localizes to the centriolar distal lumen in human cells. It is required for efficient primary ciliogenesis and nucleotide excision repair (NER). Centrin 2 forms part of the xeroderma pigmentosum group C protein complex. To explore how centrin 2 contributes to these distinct processes, we mutated the four calcium-binding EF-hand domains of human centrin 2. Centrin 2 in which all four EF-hands had been mutated to ablate calcium binding (4DA mutant) was capable of supporting in vitro NER and was as effective as the wild-type protein in rescuing the UV sensitivity of centrin 2-null cells. However, we found that mutation of any of the EF-hand domains impaired primary ciliogenesis in human TERT-RPE1 cells to the same extent as deletion of centrin 2. Phenotypic analysis of the 4DA mutant revealed defects in centrosome localization, centriole satellite assembly, ciliary assembly and function and in interactions with POC5 and SFI1. These observations indicate that centrin 2 requires calcium-binding capacity for its primary ciliogenesis functions, but not for NER, and suggest that these functions require centrin 2 to be capable of forming complexes with partner proteins. This article has an associated First Person interview with the first author of the paper.

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Section: Analysis Of the Ef-hand Domains Of Centrin 2 In Centriolar Lsupporting

confidence: 88%

Differential requirements for the EF-hand domains of human centrin 2 in primary ciliogenesis and nucleotide excision repair

Khouj

Prosser

Tada

et al. 2019

Journal of Cell Science

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“…Previous evidence has suggested that centriolar satellites are required for recruiting many key regulators of centriole duplication to the centrosome. Other possible mechanisms explaining centriolar satellite dispersion at the onset of mitosis include NEK2A kinase-dependent loss of C-NAP1 from the centrosome leading to reduced centriolar satellite density (64). There may be additional mechanisms of centriolar satellite dispersion in mitosis and reorganization in G 1 that remain to be elucidated, and this is an important area of future research.…”

Section: Plk4 Phosphorylates Cep131mentioning

confidence: 99%

Polo-like kinase 4 maintains centriolar satellite integrity by phosphorylation of centrosomal protein 131 (CEP131)

Denu

Sass

Johnson

et al. 2019

Journal of Biological Chemistry

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Edited by Xiao-Fan WangThe centrosome, consisting of two centrioles surrounded by a dense network of proteins, is the microtubule-organizing center of animal cells. Polo-like kinase 4 (PLK4) is a Ser/Thr protein kinase and the master regulator of centriole duplication, but it may play additional roles in centrosome function. To identify additional proteins regulated by PLK4, we generated an RPE-1 human cell line with a genetically engineered "analog-sensitive" PLK4 AS , which genetically encodes chemical sensitivity to competitive inhibition via a bulky ATP analog. We used this transgenic line in an unbiased multiplex phosphoproteomic screen. Several hits were identified and validated as direct PLK4 substrates by in vitro kinase assays. Among them, we confirmed Ser-78 in centrosomal protein 131 (CEP131, also known as AZI1) as a direct substrate of PLK4. Using immunofluorescence microscopy, we observed that although PLK4-mediated phosphorylation of Ser-78 is dispensable for CEP131 localization, ciliogenesis, and centriole duplication, it is essential for maintaining the integrity of centriolar satellites. We also found that PLK4 inhibition or use of a nonphosphorylatable CEP131 variant results in dispersed centriolar satellites. Moreover, replacement of endogenous WT CEP131 with an S78D phosphomimetic variant promoted aggregation of centriolar satellites. We conclude that PLK4 phosphorylates CEP131 at Ser-78 to maintain centriolar satellite integrity.

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“…These properties meet the criteria for a centrosome linker component regulated by Nek2 at the G2/M transition. Indeed, subsequent experiments have revealed that cells lacking Nek2 fail to remove C-Nap1 from centrosomes upon mitotic entry (Fletcher et al, 2004 ; Mardin et al, 2010 ), while loss of C-Nap1 by antibody microinjection, RNAi-mediated depletion, gene-editing or disease-associated truncating mutations all lead to unscheduled separation of centrosomes in interphase (Mayor et al, 2000 ; Bahe et al, 2005 ; Floriot et al, 2015 ; Panic et al, 2015 ; Flanagan et al, 2017 ).…”

Section: Nek Kinases In Centrosome Disjunctionmentioning

confidence: 99%

Mitotic Regulation by NEK Kinase Networks

Fry

Bayliss

Roig

2017

Front. Cell Dev. Biol.

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Genetic studies in yeast and Drosophila led to identification of cyclin-dependent kinases (CDKs), Polo-like kinases (PLKs) and Aurora kinases as essential regulators of mitosis. These enzymes have since been found in the majority of eukaryotes and their cell cycle-related functions characterized in great detail. However, genetic studies in another fungal species, Aspergillus nidulans, identified a distinct family of protein kinases, the NEKs, that are also widely conserved and have key roles in the cell cycle, but which remain less well studied. Nevertheless, it is now clear that multiple NEK family members act in networks to regulate specific events of mitosis, including centrosome separation, spindle assembly and cytokinesis. Here, we describe our current understanding of how the NEK kinases contribute to these processes, particularly through targeted phosphorylation of proteins associated with the microtubule cytoskeleton. We also present the latest findings on molecular events that control the activation state of the NEKs and how these are revealing novel modes of enzymatic regulation relevant not only to other kinases but also to pathological mechanisms of disease.

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Centriole splitting caused by loss of the centrosomal linker protein C-NAP1 reduces centriolar satellite density and impedes centrosome amplification

Cited by 25 publications

References 62 publications

Differential requirements for the EF-hand domains of human centrin 2 in primary ciliogenesis and nucleotide excision repair

Differential requirements for the EF-hand domains of human centrin 2 in primary ciliogenesis and nucleotide excision repair

Polo-like kinase 4 maintains centriolar satellite integrity by phosphorylation of centrosomal protein 131 (CEP131)

Mitotic Regulation by NEK Kinase Networks

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