Centriole-less pericentriolar material serves as a microtubule organizing center at the base of C. elegans sensory cilia

Magescas, Jérémy; Eskinazi, Sani; Tran, Michael V.; Feldman, Jessica L.

doi:10.1016/j.cub.2021.03.022

Cited by 33 publications

(26 citation statements)

References 37 publications

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“…However, neither of these proteins localizes to the MTOC vesicles in the PVD neuron (results not shown). This is consistent with a previous study that shows that SPD-5 mainly functions at the centrosomal MTOC (Kemp et al, 2004;Magescas et al, 2021;Magescas et al, 2019). Alternatively, NOCA-2 localization to the MTOC vesicles may be mediated by dynein.…”

Section: Noca-2 (Ninein) Acts At the Mtoc Vesiclessupporting

confidence: 93%

PTRN-1/CAMSAP and NOCA-2/NINEIN are required for microtubule polarity in Caenorhabditis elegans dendrites

Beem

Hoogenraad

et al. 2021

Preprint

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The neuronal microtubule cytoskeleton is key to establish axon-dendrite polarity. Dendrites are characterized by the presence of minus-end out microtubules, however the mechanisms that organize these microtubules minus-end out is still poorly understood. Here, we characterized the role of two microtubule minus-end related proteins in this process in Caenorhabditis elegans, the microtubule minus-end stabilizing protein CAMSAP (PTRN-1) and a NINEIN homologue (NOCA-2). We found that CAMSAP and NINEIN function in parallel to mediate microtubule organization in dendrites. During dendrite outgrowth, RAB-11 positive vesicles localized to the dendrite tip function as a microtubule organizing center (MTOC) to nucleate microtubules. In the absence of either CAMSAP or NINEIN, we observed a low penetrance MTOC vesicles mis-localization to the cell body, and a nearly fully penetrant phenotype in double mutant animals. This suggests that both proteins are important for localizing the MTOC vesicles to the growing dendrite tip to organize microtubules minus-end out. Whereas NINEIN localizes to the MTOC vesicles where it is important for the recruitment of the microtubule nucleator γ-tubulin, CAMSAP localizes around the MTOC vesicles and is co-translocated forward with the MTOC vesicles upon dendritic growth. Together, these results indicate that microtubule nucleation from the MTOC vesicles and microtubule stabilization are both important to localize the MTOC vesicles distally to organize dendritic microtubules minus-end out.

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Section: Noca-2 (Ninein) Acts At the Mtoc Vesiclessupporting

confidence: 93%

PTRN-1/CAMSAP and NOCA-2/NINEIN are required for microtubule polarity in Caenorhabditis elegans dendrites

Beem

Hoogenraad

et al. 2021

Preprint

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“…S3). While the ZF degradation system does not create a null line, it has been used to robustly deplete ZF-tagged proteins (Abrams and Nance, 2021; Liang et al, 2020; Magescas et al, 2021; Sallee et al, 2021; Sallee et al, 2018; Sanchez et al, 2021) and we see strong depletion.…”

Section: Methodsmentioning

confidence: 69%

Separable mechanisms drive local and global polarity establishment in the C. elegans intestinal epithelium

Pickett

Sallee

et al. 2021

Preprint

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Apico-basolateral polarization is essential for epithelial cells to function as selective barriers and transporters, and to provide mechanical resiliency to organs. Epithelial polarity is established locally, within individual cells to establish distinct apical, junctional, and basolateral domains, and globally, within a tissue where cells coordinately orient their apico-basolateral axes. Using live imaging of endogenously tagged proteins and tissue specific protein depletion in the C. elegans embryonic intestine, we found that local and global polarity establishment are temporally and genetically separable. Local polarity is initiated prior to global polarity and is robust to perturbation. PAR-3 is required for global polarization across the intestine but is not required for local polarity establishment as small groups of cells are able to correctly establish polarized domains in PAR-3 depleted intestines in an HMR-1/E-cadherin dependent manner. Despite belonging to the same apical protein complex, we additionally find that PAR-3 and PKC-3/aPKC have distinct roles in the establishment and maintenance of local and global polarity. Together, our results indicate that different mechanisms are required for local and global polarity establishment in vivo.SUMMARY STATEMENTLive-imaging and intestine specific protein depletion reveal that apico-basolateral polarity establishment can be temporally and genetically separated at the local level of individual cells and globally, across a tissue.

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“…Centrosomes and centrosome-related structures in C. elegans. Centrosome assembly and function has been primarily examined in the early embryo, although some limited work has also been performed on the acentriolar centrosome at the ciliary base of sensory neurons (Garbrecht et al, 2021;Magescas et al, 2021) and the non-centrosomal microtubule organizing center in the intestine (Feldman & Priess, 2012;Sanchez et al, 2021). Blast cells are two pairs of cells in the L1 larva that will give rise to the somatic gonad and germline in the adult worm.…”

Section: Mass Spectrometry Database Search and Analysismentioning

confidence: 99%

“…Recent work identifying further centriolar and pericentriolar material components whose depletion phenotypes are somewhat more subtle (Erpf et al, 2019;Sugioka et al, 2017;von Tobel et al, 2014) suggests this is not necessarily the case. Furthermore, while most of the work in C. elegans has focused on the early embryo, the available data for the acentriolar centrosome at the ciliary base of sensory neurons (Garbrecht et al, 2021;Magescas et al, 2021) and the non-centrosomal microtubule organizing center in the intestine (Feldman & Priess, 2012;Sanchez et al, 2021) (Fig. 1A) hints at tissue-dependent differences in centrosome composition and assembly mechanisms in somatic tissues of the worm that as yet remain little explored.…”

Section: Introductionmentioning

confidence: 99%

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.

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Centriole-less pericentriolar material serves as a microtubule organizing center at the base of C. elegans sensory cilia

Cited by 33 publications

References 37 publications

PTRN-1/CAMSAP and NOCA-2/NINEIN are required for microtubule polarity in Caenorhabditis elegans dendrites

PTRN-1/CAMSAP and NOCA-2/NINEIN are required for microtubule polarity in Caenorhabditis elegans dendrites

Separable mechanisms drive local and global polarity establishment in the C. elegans intestinal epithelium

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

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