Centriole distal-end proteins CP110 and Cep97 influence centriole cartwheel growth at the proximal-end

Aydogan, Mustafa G.; Hankins, Laura E; Steinacker, Thomas L.; Mofatteh, Mohammad; Saurya, Saroj; Wainman, Alan; Wong, Siu‐Shing; Lü, Xin; Zhou, Felix; Raff, Jordan W.

doi:10.1101/2021.07.08.451650

Cited by 5 publications

(2 citation statements)

References 48 publications

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“…However, it is striking to note that CCEP-97 is first detectable at centrioles as the cell cycle slows in late-stage embryos and strongly accumulates at centrioles in gonad precursor cells, cells which as previously noted are arrested for extended periods of time in interphase. These findings are consistent with recent reports of cell cycle-entrained oscillations of Cep97 levels regulating centriole growth in Drosophila [ 74 ].…”

Section: Resultssupporting

confidence: 94%

“…Other proteins are only detected at later developmental stages, such as CCEP-97. Its appearance at centrioles in late-stage embryos may reflect a slowing of the cell cycle allowing its accumulation during the more extended interphase [ 74 ], and be connected to the emergence of centriolar doublet microtubules at approximately the same time [ 51 , 80 ]. Finally, ciliated sensory neurons present a unique set of SPD-5 proximity interactors, including CHE-10/rootletin.…”

Section: Discussionmentioning

confidence: 99%

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A modified TurboID approach identifies tissue-specific centriolar components in C. elegans

et al. 2022

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Proximity-dependent labeling approaches such as BioID have been a great boon to studies of protein-protein interactions in the context of cytoskeletal structures such as centrosomes which are poorly amenable to traditional biochemical approaches like immunoprecipitation and tandem affinity purification. Yet, these methods have so far not been applied extensively to invertebrate experimental models such as C. elegans given the long labeling times required for the original promiscuous biotin ligase variant BirA*. Here, we show that the recently developed variant TurboID successfully probes the interactomes of both stably associated (SPD-5) and dynamically localized (PLK-1) centrosomal components. We further develop an indirect proximity labeling method employing a GFP nanobody-TurboID fusion, which allows the identification of protein interactors in a tissue-specific manner in the context of the whole animal. Critically, this approach utilizes available endogenous GFP fusions, avoiding the need to generate multiple additional strains for each target protein and the potential complications associated with overexpressing the protein from transgenes. Using this method, we identify homologs of two highly conserved centriolar components, Cep97 and BLD10/Cep135, which are present in various somatic tissues of the worm. Surprisingly, neither protein is expressed in early embryos, likely explaining why these proteins have escaped attention until now. Our work expands the experimental repertoire for C. elegans and opens the door for further studies of tissue-specific variation in centrosome architecture.

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Section: Resultssupporting

confidence: 94%

Section: Discussionmentioning

confidence: 99%

A modified TurboID approach identifies tissue-specific centriolar components in C. elegans

et al. 2022

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Self-assembly of pericentriolar material in interphase cells lacking centrioles

Chen

Iwanski

et al. 2021

Preprint

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The major microtubule-organizing center (MTOC) in animal cells, the centrosome, comprises a pair of centrioles surrounded by pericentriolar material (PCM), which nucleates and anchors microtubules. Centrosome assembly depends on the interactions of PCM with centrioles, PCM self-association and dynein-mediated transport. Here, we show that if centrioles are lost due to PLK4 depletion or inhibition, PCM still forms a single centrally located MTOC when non-centrosomal microtubule minus end organization pathways are disabled. Acentriolar MTOC assembly depends on dynein-driven coalescence of PCM clusters with attached microtubule minus ends and requires γ-tubulin, pericentrin, CDK5RAP2 and ninein, but not NEDD1, CEP152 or CEP192. PCM self-assembly is inhibited by AKAP450-dependent PCM recruitment to the Golgi and by CAMSAP2-mediated microtubule minus end stabilization. However, if CAMSAP2 is linked to a minus-end-directed motor, a single MTOC containing PCM components can still form, and its organization depends on the presence of pericentrin. Our results reveal that the formation of a single central MTOC in interphase mammalian cells is not strictly centriole dependent but can be driven by self-organization of PCM and microtubule minus ends.

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A Modified TurboID Approach Identifies Tissue-Specific Centriolar Components InC. elegans

Holzer

Rumpf-Kienzl

Falk

et al. 2021

Preprint

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Proximity-dependent labeling approaches such as BioID have been a great boon to studies of protein-protein interactions in the context of cytoskeletal structures such as centrosomes which are poorly amenable to traditional biochemical approaches like immunoprecipitation and tandem affinity purification. Yet, these methods have so far not been applied extensively to invertebrate experimental models such as C. elegans given the long labeling times required for the original promiscuous biotin ligase variant BirA*. Here, we show that the recently developed variant TurboID successfully probes the interactomes of both stably associated (SPD-5) and dynamically localized (PLK-1) centrosomal components. We further develop an indirect proximity labeling method employing a GFP nanobody- TurboID fusion, which allows the identification of protein interactors in a tissue-specific manner in the context of the whole animal. Critically, this approach utilizes available endogenous GFP fusions, avoiding the need to generate multiple additional strains for each target protein and the potential complications associated with overexpressing the protein from transgenes. Using this method, we identify homologs of two highly conserved centriolar components, Cep97 and Bld10/Cep135, which are present in various somatic tissues of the worm. Surprisingly, neither protein is expressed in early embryos, likely explaining why these proteins have escaped attention until now. Our work expands the experimental repertoire for C. elegans and opens the door for further studies of tissue-specific variation in centrosome architecture.

show abstract

Centriole distal-end proteins CP110 and Cep97 influence centriole cartwheel growth at the proximal-end

Cited by 5 publications

References 48 publications

A modified TurboID approach identifies tissue-specific centriolar components in C. elegans

A modified TurboID approach identifies tissue-specific centriolar components in C. elegans

Self-assembly of pericentriolar material in interphase cells lacking centrioles

A Modified TurboID Approach Identifies Tissue-Specific Centriolar Components InC. elegans

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