1. Cell Biology

Myristoylated Neuronal Calcium Sensor-1 captures the ciliary vesicle at distal appendages

  1. Tomoharu Kanie  Is a corresponding author
  2. Roy Ng
  3. Keene L Abbott
  4. Niaj Mohammad Tanvir
  5. Esben Lorentzen
  6. Olaf Pongs
  7. Peter K Jackson  Is a corresponding author
  1. University of Oklahoma Health Sciences Center, United States
  2. Stanford University, United States
  3. Aarhus University, Denmark
  4. Saarland University, Germany
https://doi.org/10.7554/eLife.85998
Share this article
Cite this article
  1. Tomoharu Kanie
  2. Roy Ng
  3. Keene L Abbott
  4. Niaj Mohammad Tanvir
  5. Esben Lorentzen
  6. Olaf Pongs
  7. Peter K Jackson
(2025)
Myristoylated Neuronal Calcium Sensor-1 captures the ciliary vesicle at distal appendages
eLife 14:e85998.
https://doi.org/10.7554/eLife.85998
  2. Download BibTeX
  3. Download .RIS

Abstract

The primary cilium is a microtubule-based organelle that cycles through assembly and disassembly. In many cell types, formation of the cilium is initiated by recruitment of ciliary vesicles to the distal appendage of the mother centriole. However, the distal appendage mechanism that directly captures ciliary vesicles is yet to be identified. In an accompanying paper, we show that the distal appendage protein, CEP89, is important for the ciliary vesicle recruitment, but not for other steps of cilium formation (Tomoharu Kanie, Love, Fisher, Gustavsson, & Jackson, 2023). The lack of a membrane binding motif in CEP89 suggests that it may indirectly recruit ciliary vesicles via another binding partner. Here, we identify Neuronal Calcium Sensor-1 (NCS1) as a stoichiometric interactor of CEP89. NCS1 localizes to the position between CEP89 and a ciliary vesicle marker, RAB34, at the distal appendage. This localization was completely abolished in CEP89 knockouts, suggesting that CEP89 recruits NCS1 to the distal appendage. Similarly to CEP89 knockouts, ciliary vesicle recruitment as well as subsequent cilium formation was perturbed in NCS1 knockout cells. The ability of NCS1 to recruit the ciliary vesicle is dependent on its myristoylation motif and NCS1 knockout cells expressing a myristoylation defective mutant failed to rescue the vesicle recruitment defect despite localizing properly to the centriole. In sum, our analysis reveals the first known mechanism for how the distal appendage recruits the ciliary vesicles.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for corresponding figures.

Article and author information

Author details

  1. Tomoharu Kanie

    Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, United States
    For correspondence
    Tomoharu-Kanie@ouhsc.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2084-1451

  2. Roy Ng

    Department of Microbiology and Immunology, Stanford University, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Keene L Abbott

    Department of Microbiology and Immunology, Stanford University, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6166-704X

  4. Niaj Mohammad Tanvir

    Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
    Competing interests
    The authors declare that no competing interests exist.

  5. Esben Lorentzen

    Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6493-7220

  6. Olaf Pongs

    Institute for Physiology, Saarland University, Saarbrücken, Germany
    Competing interests
    The authors declare that no competing interests exist.

  7. Peter K Jackson

    Department of Microbiology and Immunology, Stanford University, Stanford, United States
    For correspondence
    pjackson@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.

Funding

National Institute of General Medical Sciences (P20GM103447)

  • Tomoharu Kanie

National Institute of General Medical Sciences (1R35GM151013)

  • Tomoharu Kanie

National Institute of General Medical Sciences (R01GM114276)

  • Peter K Jackson

National Institute of General Medical Sciences (R01GM121565)

  • Peter K Jackson

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All mice were maintained under specific pathogen-free conditions at the Stanford animal care facility. All experiments were approved by Administrative Panel on Laboratory Animal Care at Stanford University (Institutional Animal Care and Use Committee protocol number: 28556) and were performed in strict accordance with their guidelines.

Copyright

© 2025, Kanie et al.

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 389
    views
  • 91
    downloads
  • 1
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Tomoharu Kanie
  2. Roy Ng
  3. Keene L Abbott
  4. Niaj Mohammad Tanvir
  5. Esben Lorentzen
  6. Olaf Pongs
  7. Peter K Jackson
(2025)
Myristoylated Neuronal Calcium Sensor-1 captures the ciliary vesicle at distal appendages
eLife 14:e85998.
https://doi.org/10.7554/eLife.85998

Share this article

https://doi.org/10.7554/eLife.85998

Categories and tags

Research organisms

Further reading

    1. Cell Biology

    A hierarchical pathway for assembly of the distal appendages that organize primary cilia

    Tomoharu Kanie, Beibei Liu ... Peter K Jackson
    Research Article Updated

    Distal appendages are ninefold symmetric blade-like structures attached to the distal end of the mother centriole. These structures are critical for the formation of the primary cilium, by regulating at least four critical steps: preciliary vesicle recruitment, recruitment and initiation of intraflagellar transport (IFT), and removal of CP110. While specific proteins that localize to the distal appendages have been identified, how exactly each protein functions to achieve the multiple roles of the distal appendages is poorly understood. Here, we comprehensively analyze known and newly discovered distal appendage proteins (CEP83, SCLT1, CEP164, TTBK2, FBF1, CEP89, KIZ, ANKRD26, PIDD1, LRRC45, NCS1, CEP15) for their precise localization, order of recruitment, and their roles in each step of cilia formation. Using CRISPR-Cas9 knockouts, we show that the order of the recruitment of the distal appendage proteins is highly interconnected and a more complex hierarchy. Our analysis highlights two protein modules, CEP83-SCLT1 and CEP164-TTBK2, as critical for structural assembly of distal appendages. Functional assays revealed that CEP89 selectively functions in the RAB34+ vesicle recruitment, while deletion of the integral components, CEP83-SCLT1-CEP164-TTBK2, severely compromised all four steps of cilium formation. Collectively, our analyses provide a more comprehensive view of the organization and the function of the distal appendage, paving the way for molecular understanding of ciliary assembly.

    1. Cell Biology
    2. Medicine

    Disruption of the novel nested gene Aff3ir mediates disturbed flow-induced atherosclerosis in mice

    Shuo He, Lei Huang ... Jinlong He
    Research Article

    Disturbed shear stress-induced endothelial atherogenic responses are pivotal in the initiation and progression of atherosclerosis, contributing to the uneven distribution of atherosclerotic lesions. This study investigates the role of Aff3ir-ORF2, a novel nested gene variant, in disturbed flow-induced endothelial cell activation and atherosclerosis. We demonstrate that disturbed shear stress significantly reduces Aff3ir-ORF2 expression in athero-prone regions. Using three distinct mouse models with manipulated Aff3ir-ORF2 expression, we demonstrate that Aff3ir-ORF2 exerts potent anti-inflammatory and anti-atherosclerotic effects in Apoe-/- mice. RNA sequencing revealed that interferon regulatory factor 5 (Irf5), a key regulator of inflammatory processes, mediates inflammatory responses associated with Aff3ir-ORF2 deficiency. Aff3ir-ORF2 interacts with Irf5, promoting its retention in the cytoplasm, thereby inhibiting the Irf5-dependent inflammatory pathways. Notably, Irf5 knockdown in Aff3ir-ORF2 deficient mice almost completely rescues the aggravated atherosclerotic phenotype. Moreover, endothelial-specific Aff3ir-ORF2 supplementation using the CRISPR/Cas9 system significantly ameliorated endothelial activation and atherosclerosis. These findings elucidate a novel role for Aff3ir-ORF2 in mitigating endothelial inflammation and atherosclerosis by acting as an inhibitor of Irf5, highlighting its potential as a valuable therapeutic approach for treating atherosclerosis.

    1. Cell Biology
    2. Developmental Biology

    Cell Signaling: Not all dimers are equal

    Jeet H Patel, Mary C Mullins
    Insight

    Disease-causing mutations in the signaling protein BMP4 impair its secretion, but only when it is made as a homodimer.