Stromule extension along microtubules coordinated with actin-mediated anchoring guides perinuclear chloroplast movement during innate immunity

Abstract

Dynamic tubular extensions from chloroplasts called stromules have recently been shown to connect with nuclei and function during innate immunity. We demonstrate that stromules extend along microtubules (MTs) and MT organization directly affects stromule dynamics since stabilization of MTs chemically or genetically increases stromule numbers and length. Although actin filaments (AFs) are not required for stromule extension, they provide anchor points for stromules. Interestingly, there is a strong correlation between the direction of stromules from chloroplasts and the direction of chloroplast movement. Stromule-directed chloroplast movement was observed in steady-state conditions without immune induction, suggesting it is a general function of stromules in epidermal cells. Our results show that MTs and AFs may facilitate perinuclear clustering of chloroplasts during an innate immune response. We propose a model in which stromules extend along MTs and connect to AF anchor points surrounding nuclei, facilitating stromule-directed movement of chloroplasts to nuclei during innate immunity.

Article and author information

Author details

  1. Amutha Sampath Kumar

    Delaware Biotechnology Insititute, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Eunsook Park

    Department of Plant and The Genome Center, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2984-3039
  3. Alexander Nedo

    Delaware Biotechnology Insititute, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Ali Alqarni

    Delaware Biotechnology Insititute, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Li Ren

    Department of Plant and Soil Sciences, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Kyle Hoban

    Delaware Biotechnology Institute, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Shannon Modla

    Delaware Biotechnology Institute, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. John H McDonald

    Department of Biological Sciences, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Chandra Kambhamettu

    Department of Plant and Soil Sciences, University of Delaware, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Savithramma P Dinesh-Kumar

    Department of Plant and The Genome Center, University of California, Davis, Davis, United States
    For correspondence
    spdineshkumar@ucdavis.edu
    Competing interests
    The authors declare that no competing interests exist.
  11. Jeffrey Lewis Caplan

    Delaware Biotechnology Institute, University of Delaware, Newark, United States
    For correspondence
    jcaplan@udel.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3991-0912

Funding

National Institutes of Health (R01 GM097587)

  • Savithramma P Dinesh-Kumar
  • Jeffrey Lewis Caplan

National Institutes of Health (P20 GM103446)

  • Jeffrey Lewis Caplan

National Institutes of Health (S10 OD016361)

  • Jeffrey Lewis Caplan

National Institutes of Health (S10 RR027273)

  • Jeffrey Lewis Caplan

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

Copyright

© 2018, Kumar et al.

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

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  1. Amutha Sampath Kumar
  2. Eunsook Park
  3. Alexander Nedo
  4. Ali Alqarni
  5. Li Ren
  6. Kyle Hoban
  7. Shannon Modla
  8. John H McDonald
  9. Chandra Kambhamettu
  10. Savithramma P Dinesh-Kumar
  11. Jeffrey Lewis Caplan
(2018)
Stromule extension along microtubules coordinated with actin-mediated anchoring guides perinuclear chloroplast movement during innate immunity
eLife 7:e23625.
https://doi.org/10.7554/eLife.23625

Share this article

https://doi.org/10.7554/eLife.23625

Further reading

    1. Cell Biology
    Rachel Pudlowski, Lingyi Xu ... Jennifer T Wang
    Research Advance

    Centrioles have a unique, conserved architecture formed by three linked, ‘triplet’, microtubules arranged in ninefold symmetry. The mechanisms by which these triplet microtubules are formed remain unclear but likely involve the noncanonical tubulins delta-tubulin and epsilon-tubulin. Previously, we found that human cells lacking delta-tubulin or epsilon-tubulin form abnormal centrioles, characterized by an absence of triplet microtubules, lack of central core protein POC5, and a futile cycle of centriole formation and disintegration (Wang et al., 2017). Here, we show that human cells lacking either TEDC1 or TEDC2 have similar abnormalities. Using ultrastructure expansion microscopy, we observed that mutant centrioles elongate to the same length as control centrioles in G2 phase and fail to recruit central core scaffold proteins. Remarkably, mutant centrioles also have an expanded proximal region. During mitosis, these mutant centrioles further elongate before fragmenting and disintegrating. All four proteins physically interact and TEDC1 and TEDC2 can form a subcomplex in the absence of the tubulins, supporting an AlphaFold Multimer model of the tetramer. TEDC1 and TEDC2 localize to centrosomes and are mutually dependent on each other and on delta-tubulin and epsilon-tubulin for localization. Our results demonstrate that delta-tubulin, epsilon-tubulin, TEDC1, and TEDC2 function together to promote robust centriole architecture, laying the foundation for future studies on the mechanisms underlying the assembly of triplet microtubules and their interactions with centriole structure.

    1. Cell Biology
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    Pengbo Chen, Bo Li ... Xinfeng Zheng
    Research Article

    Background:

    It has been reported that loss of PCBP2 led to increased reactive oxygen species (ROS) production and accelerated cell aging. Knockdown of PCBP2 in HCT116 cells leads to significant downregulation of fibroblast growth factor 2 (FGF2). Here, we tried to elucidate the intrinsic factors and potential mechanisms of bone marrow mesenchymal stromal cells (BMSCs) aging from the interactions among PCBP2, ROS, and FGF2.

    Methods:

    Unlabeled quantitative proteomics were performed to show differentially expressed proteins in the replicative senescent human bone marrow mesenchymal stromal cells (RS-hBMSCs). ROS and FGF2 were detected in the loss-and-gain cell function experiments of PCBP2. The functional recovery experiments were performed to verify whether PCBP2 regulates cell function through ROS/FGF2-dependent ways.

    Results:

    PCBP2 expression was significantly lower in P10-hBMSCs. Knocking down the expression of PCBP2 inhibited the proliferation while accentuated the apoptosis and cell arrest of RS-hBMSCs. PCBP2 silence could increase the production of ROS. On the contrary, overexpression of PCBP2 increased the viability of both P3-hBMSCs and P10-hBMSCs significantly. Meanwhile, overexpression of PCBP2 led to significantly reduced expression of FGF2. Overexpression of FGF2 significantly offset the effect of PCBP2 overexpression in P10-hBMSCs, leading to decreased cell proliferation, increased apoptosis, and reduced G0/G1 phase ratio of the cells.

    Conclusions:

    This study initially elucidates that PCBP2 as an intrinsic aging factor regulates the replicative senescence of hBMSCs through the ROS-FGF2 signaling axis.

    Funding:

    This study was supported by the National Natural Science Foundation of China (82172474).