Microtubule re-organization during female meiosis in C elegans
- Technische Universitaet Dresden, Germany
- University of California, Santa Barbara, United States
- University of Virginia, United States
- Zuse Institute Berlin, Germany
- University of Alberta, Canada
- Flatiron Institute, United States
Abstract
The female meiotic spindles of most animals are acentrosomal and undergo striking morphological changes while transitioning from metaphase to anaphase. The ultra-structure of acentrosomal spindles, and how changes to this structure correlate with such dramatic spindle rearrangements remains largely unknown. To address this, we applied light microscopy, large-scale electron tomography and mathematical modeling of female meiotic C. elegans spindles undergoing the transition from metaphase to anaphase. Combining these approaches, we find that meiotic spindles are dynamic arrays of short microtubules that turn over on second time scales. The results show that the transition from metaphase to anaphase correlates with an increase in the number of microtubules and a decrease in their average length. Detailed analysis of the tomographic data revealed that the length of microtubules changes significantly during the metaphase-to-anaphase transition. This effect is most pronounced for those microtubules located within 150 nm of the chromosome surface. To understand the mechanisms that drive this transition, we developed a mathematical model for the microtubule length distribution that considers microtubule growth, catastrophe, and severing. Using Bayesian inference to compare model predictions and data, we find that microtubule turn-over is the major driver of the observed large-scale reorganizations. Our data suggest that in metaphase only a minor fraction of microtubules, those that are closest to the chromosomes, are severed. The large majority of microtubules, which are not in close contact with chromosomes, do not undergo severing. Instead, their length distribution is fully explained by growth and catastrophe alone. In anaphase, even microtubules close to the chromosomes show no signs of cutting. This suggests that the most prominent drivers of spindle rearrangements from metaphase to anaphase are changes in nucleation and catastrophe rate. In addition, we provide evidence that microtubule severing is dependent on the presence of katanin.
Data availability
Electron microscopy models of microtubules and chromosome surfaces will be made available on Dryad under doi.org/10.5061/dryad.x3ffbg7k5. Example data and analysis code is available at https://github.com/SebastianFuerthauer/SpindleRerrangement
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C. elegans meiotic spindlesDryad Digital Repository, doi.org/10.5061/dryad.x3ffbg7k5.
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Meiosis I spindles of Metaphase, early Anaphase and Anaphasehttps://www.cell.com/current-biology/fulltext/S0960-9822(18)30911-4?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS0960982218309114%3Fshowall%3Dtrue.
Article and author information
Author details
Funding
Deutsche Forschungsgemeinschaft (MU 1423/3-1)
- Ina Lantzsch
Deutsche Forschungsgemeinschaft (MU 1423/3-2)
- Ina Lantzsch
Deutsche Forschungsgemeinschaft (MU 1423/8-1)
- Erik Szentgyoergyi
Technische Universität Darmstadt (Frauenhabilitation)
- Stefanie Redemann
Natural Sciences and Engineering Research Council of Canada
- Martin Srayko
National Science Foundation (DMR-0820484)
- Che-Hang Yu
National Science Foundation (NeuroNex #1934288)
- Che-Hang Yu
National Institutes of Health (1R01GM104976-01)
- Che-Hang Yu
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Lantzsch 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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Microtubule re-organization during female meiosis in C elegans
eLife 10:e58903.
https://doi.org/10.7554/eLife.58903
Further reading
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- Cell Biology
Polynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment to the gapped DNA structure found in single-strand DNA (ssDNA) nicks and/or gaps between Okazaki fragments (OFs) during DNA replication. T118A (alanine)-substituted PNKP-expressing cells exhibited an accumulation of ssDNA gaps in S phase and accelerated replication fork progression. Furthermore, PNKP is involved in poly (ADP-ribose) polymerase 1 (PARP1)-dependent replication gap filling as part of a backup pathway in the absence of OFs ligation. Altogether, our data suggest that CDK-mediated PNKP phosphorylation at T118 is important for its recruitment to ssDNA gaps to proceed with OFs ligation and its backup repairs via the gap-filling pathway to maintain genome stability.
