Centriolar satellites are acentriolar assemblies of centrosomal proteins

Quarantotti, Valentina; Chen, Jia‐Xuan; Tischer, Julia; Tejedo, Carmen González; Papachristou, Evaggelia K.; D’Santos, Clive S.; Kilmartin, John V.; Miller, Martin L.; Gergely, Fanni

doi:10.15252/embj.2018101082

Cited by 68 publications

(113 citation statements)

References 82 publications

(147 reference statements)

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“…Notably, fibrogranular material in MCCs was shown to contain the protein PCM1 39 , which is also required to scaffold the assembly of the 'pericentriolar satellites' that are found in non-MCCs. It is therefore possible that the fibrogranular material is functionally equivalent to the pericentriolar satellites of non-MCCs [40][41][42][43] . Additional studies will be required to identify the minimal components involved in centriole assembly and number control, and to define the role of DEUP1 and the deuterosome in MCCs.…”

Section: Discussionmentioning

confidence: 99%

Massive centriole production can occur in the absence of deuterosomes in multiciliated cells

et al. 2019

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ulticiliated cells (MCCs) contain tens of motile cilia that beat to drive fluid flow across epithelial surfaces. Multiciliated cells are present in the respiratory tract, brain ventricles and reproductive systems. Defects in motile-cilia formation or beating lead to the development of hydrocephaly, lethal respiratory symptoms and fertility defects 1-4. A centriole, or basal body, serves as a template for the cilium axoneme. Centriole duplication is tightly controlled in cycling cells so that a single new procentriole forms adjacent to each of the two parent centrioles 5. However, MCC progenitors with two parent centrioles produce tens to hundreds of additional new centrioles to nucleate multiple motile cilia 1. Steric constraints imposed by the 'centriolar' pathway seem to restrict the number of procentrioles that can be nucleated by the parent centrioles. Centriole amplification is therefore thought to rely on the assembly of dozens of MCCspecific organelles called deuterosomes, which each nucleate tens of procentrioles 6-14. Deuterosomes are assembled during centriole amplification and support the growth of approximately 90% of the procentrioles formed in mammalian MCCs 12,14. Deuterosomes have been proposed to be nucleated from the younger parent centriole 12 but can form spontaneously in a cloud of pericentriolar material (PCM) in MCCs depleted of the parent centrioles 15-17. Many of the proteins required for centriole formation in MCCs are common to centriole duplication 11-15,18-23. However, DEUP1 (CCDC67, alternate gene name) has been identified as a deuterosome-specific protein that arose from a gene-duplication event of the centriolar gene Cep63. Recent data suggest that Deup1 evolved to enable the formation of deuterosomes and the generation of large numbers of centrioles 14. In this manuscript, we interrogate the function of the deuterosome in MCCs from mouse and from Xenopus laevis. Surprisingly, our findings reveal that deuterosomes are dispensable for centriole amplification and multiciliogenesis both in vitro and in vivo. Moreover, we show that neither deuterosomes nor parent centrioles are required for MCCs to amplify the correct number of centrioles. These findings raise new questions about the evolutionary role of deuterosome during multiciliogenesis and the mechanisms regulating centriole number in MCCs. Results Generation of a Deup1-knockout mouse. To examine the role of the deuterosome in multiciliogenesis we created a Deup1-knockout mouse by replacing a region from within exon 2 to within exon 7 of the Deup1 gene with a LacZ reporter (Extended Data Fig. 1a). Reverse Transcription-quantitative PCR on brain and testes samples showed that the messenger RNA levels of Deup1 were reduced by at least tenfold in Deup1-knockout compared with control mice (Extended Data Fig. 1b,c). To examine the process of multiciliogenesis in Deup1 −/− cells, we utilized in vitro cultures of mouse tracheal epithelial cells (mTECs) or ependymal cells 24,25. Consistent with the absence of Deup1 mRNA, DEUP1 foci were a...

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Section: Discussionmentioning

confidence: 99%

Massive centriole production can occur in the absence of deuterosomes in multiciliated cells

et al. 2019

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“…We also noted that centrin 2 loss or calcium-binding deficiency impacted on centriolar satellite aggregation, as measured by PCM1 signal. Centriolar satellites have been suggested to act as regulators of centrosome duplication by controlling availability of centrosome components (Dammermann and Merdes, 2002;Kubo and Tsukita, 2003;Firat-Karalar et al, 2014;Tollenaere et al, 2015) and more recent data have similarly implicated them in ciliogenesis (Odabasi et al, 2019;Quarantotti et al, 2019). The reduction of ciliogenesis in cells deficient in centrin 2 or expressing an EF-hand mutant might thus reflect deficient aggregation of centrosome regulators in centriolar satellites.…”

Section: Discussionmentioning

confidence: 99%

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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“…Centriolar satellites (hereafter satellites) are 70-100-nm membraneless electron-dense granules that localize and move around centrosomes and cilia [1]. Satellite assembly is scaffolded by the large coiled-coil protein pericentriolar material 1 (PCM1), which physically interacts with many known and putative centrosome proteins [2,3]. Satellite resident proteins function in a wide range of cellular processes, including cilium assembly, ciliary transport, centriole duplication, mitotic regulation, and microtubule dynamics and organization, and include proteins mutated in developmental and neuronal disorders [4,5].…”

Section: Introductionmentioning

confidence: 99%

Acute inhibition of centriolar satellite function and positioning reveals their functions at the primary cilium

et al. 2020

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Centriolar satellites are dynamic, membraneless granules composed of over 200 proteins. They store, modify, and traffic centrosome and primary cilium proteins, and help to regulate both the biogenesis and some functions of centrosomes and cilium. In most cell types, satellites cluster around the perinuclear centrosome, but their integrity and cellular distribution are dynamically remodeled in response to different stimuli, such as cell cycle cues. Dissecting the specific and temporal functions and mechanisms of satellites and how these are influenced by their cellular positioning and dynamics has been challenging using genetic approaches, particularly in ciliated and proliferating cells. To address this, we developed a chemical-based trafficking assay to rapidly and efficiently redistribute satellites to either the cell periphery or center, and fuse them into stable clusters in a temporally controlled way. Induced satellite clustering at either the periphery or center resulted in antagonistic changes in the pericentrosomal levels of a subset of proteins, revealing a direct and selective role for their positioning in protein targeting and sequestration. Systematic analysis of the interactome of peripheral satellite clusters revealed enrichment of proteins implicated in cilium biogenesis and mitosis. Importantly, induction of peripheral satellite targeting in ciliated cells revealed a function for satellites not just for efficient cilium assembly but also in the maintenance of steady-state cilia and in cilia disassembly by regulating the structural integrity of the ciliary axoneme. Finally, perturbing satellite distribution and dynamics inhibited their mitotic dissolution, and mitotic progression was perturbed only in cells with centrosomal satellite clustering. Collectively, our results for the first time showed a direct link between satellite functions and their pericentrosomal clustering, suggested new mechanisms underlying satellite functions during cilium assembly, and provided a new tool for probing temporal satellite functions in different contexts

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Centriolar satellites are acentriolar assemblies of centrosomal proteins

Cited by 68 publications

References 82 publications

Massive centriole production can occur in the absence of deuterosomes in multiciliated cells

Massive centriole production can occur in the absence of deuterosomes in multiciliated cells

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

Acute inhibition of centriolar satellite function and positioning reveals their functions at the primary cilium

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