PLK4 drives centriole amplification and apical surface area expansion in multiciliated cells

  1. Gina M LoMastro
  2. Chelsea G Drown
  3. Aubrey L Maryniak
  4. Cayla E Jewett
  5. Margaret A Strong
  6. Andrew Jon Holland

  • Curated by eLife

    eLife logo

    Evaluation Summary:

    PLK4 is the master regulator of centriole biogenesis, but whether it is also key for centriole amplification during differentiation of multiciliated cells (MCCs) has been questioned based on PLK4 chemical inhibition. Here, using mouse models engineered to lack PLK4 or PLK4 activity, LoMastro et al provide very compelling evidence that PLK4 and its activity are essential for centriole amplification in MCCs. Moreover, they show that centriole amplification in MCCs drives expansion of their apical surface. The findings will be of interest to cell biologists and experts interested in multi-ciliogenesis-related pathologies.

    (This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

Multiciliated cells (MCCs) are terminally differentiated epithelia that assemble multiple motile cilia used to promote fluid flow. To template these cilia, MCCs dramatically expand their centriole content during a process known as centriole amplification. In cycling cells, the master regulator of centriole assembly Polo-like kinase 4 (PLK4) is essential for centriole duplication; however recent work has questioned the role of PLK4 in centriole assembly in MCCs. To address this discrepancy, we created genetically engineered mouse models and demonstrated that both PLK4 protein and kinase activity are critical for centriole amplification in MCCs. Tracheal epithelial cells that fail centriole amplification accumulate large assemblies of centriole proteins and do not undergo apical surface area expansion. These results show that the initial stages of centriole assembly are conserved between cycling cells and MCCs and suggest that centriole amplification and surface area expansion are coordinated events.

Article activity feed

  1. Version published to 10.7554/elife.80643 on eLife
  2. eLife

    Evaluation Summary:

    PLK4 is the master regulator of centriole biogenesis, but whether it is also key for centriole amplification during differentiation of multiciliated cells (MCCs) has been questioned based on PLK4 chemical inhibition. Here, using mouse models engineered to lack PLK4 or PLK4 activity, LoMastro et al provide very compelling evidence that PLK4 and its activity are essential for centriole amplification in MCCs. Moreover, they show that centriole amplification in MCCs drives expansion of their apical surface. The findings will be of interest to cell biologists and experts interested in multi-ciliogenesis-related pathologies.

    (This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

  3. eLife

    Reviewer #1 (Public Review):

    The manuscript by LoMastro et al. investigates whether Plk4, the master regulator of centriole biogenesis in cycling cells, has a similar role during the differentiation of multi-ciliated cells, which produce tens to hundreds of centrioles during multi-ciliogenesis. Contrasting previous work that did not find an important role for Plk4 in this process based on chemical inhibition, the authors in the current study use genetic approaches and mouse models to show that Plk4 and its kinase activity are essential for centriole amplification and multi-ciliogenesis in two different multi-ciliated cell types in vitro and in vivo. In addition, they show that centriole amplification drives cell surface area expansion.

    The study addresses an important question regarding the role of Plk4 in centriole amplification during multi-ciliogenesis. It convincingly establishes that contrary to previous findings, the Plk4-dependent control of centriole biogenesis that is well-established in cycling cells is conserved also during differentiation of multi-ciliated cells. The presented data is of very high quality, phenotypes are well described and quantified, the conclusions are clear, and obtained in both in vitro and in vivo models. The authors also test chemical inhibition of Plk4 as used in previous work and show that the lack of a strong phenotypes under these conditions is likely due to incomplete Plk4 inhibition.

  4. eLife

    Reviewer #2 (Public Review):

    PLK4 is the master regulator of centriole assembly in cycling cells, but whether it is also key for centriole amplification during differentiation of multiciliated cells (MCCs) had been questioned. In this manuscript, LoMastro et al provide very compelling evidence that PLK4 and its kinase activity are indeed essential for centriole amplification in MCCs, and that previous failure to see these effects was due to insufficient knockdown/inhibition. LoMastro et al demonstrate this very elegantly in both ependymal and tracheal MCC mouse models, which they genetically engineer to completely lack PLK4 (KO), or to completely lack its activity (knockin of kinase-dead PLK4). These and other tools generated herein will likely be of great value in future studies addressing PLK4 and centriolar functions. Furthermore, they show that centriole amplification in MCCs drives expansion of their apical surface, an important insight to understand their differentiation.

  5. Version published to 10.1101/2022.06.06.494981v1 on bioRxiv