Drosophila kinesin-8 stabilises kinetochore-microtubule interaction
Posted on: 16 November 2018
Preprint posted on 25 October 2018
Article now published in Journal of Cell Biology at http://dx.doi.org/10.1083/jcb.201807077
Sizing up the mitotic spindle: KLP67A regulates microtubule dynamics and stabilises kinetochore-microtubule attachments in Drosophila.
Selected by Ben Craske, Gaetan Dias Mirandela, Thibault Legal and Toni McHughCategories: biochemistry, cell biology
Context:
The collective efforts of microtubule motors during cell division facilitates the organisation of the bipolar spindle, the establishment of stable kinetochore – microtubule attachments and faithful segregation of genetic material. Kinesin-8 is a widely conserved processive motor which generally localises to the kinetochore and is required for proper chromosome alignment in budding yeast and mammalian cells. However, the proposed role of Kinesin-8 motors as active microtubule depolymerases is under debate. Here, the authors investigate the motility and depolymerase activity of the sole Drosophila Kinesin-8 motor KLP67A using in vitro reconstitutions with both GMPCPP stabilised and dynamic microtubules. Additionally, they characterise the importance of KLP67A in stabilising the kinetochore-microtubule attachment and regulating spindle length in mitotic S2 cells.
Key findings:
As is characteristic of Kinesin-8 motors, the authors demonstrate that full length Drosophila KLP67A is able to processively walk along microtubules and accumulate at the plus tips in vitro. However, no detectable depolymerisation phenotype was observed for KLP67A after application to GMPCPP stabilised microtubules, in contrast to what has been characterised for S. cerevisiae Kip3 or Drosophila Kinesin-13 depolymerase, KLP10A. Despite the absence of obvious depolymerase activity upon stabilised microtubules, the localisation of KLP67A to the tips of dynamic microtubules induced a significantly higher frequency of both catastrophe and subsequent rescue. This indicates that the sole Drosophila Kinesin-8 motor is able to induce microtubule depolymerisation, but also provide a subsequent stabilising effect at the plus tip to prevent severe shrinkage events and therefore operating by a distinct mechanism to both KLP10A and Kip3.
Figure adapted from Edzuka and Goshima, 2018
In S2 cells, RNAi knockdowns of KLP67A resulted in extensive elongation of the spindle microtubules, consistent with their conclusion that Drosophila Kinesin-8 can regulate the length of dynamic microtubules in vitro. In order to investigate KLP67A’s potential function in stabilising the kinetochore – microtubule attachment, S2 cells with monopolar spindles were generated and GFP-Rod was used as a marker of kinetochore attachment status following KLP67A knockdown. Under such conditions, the authors observed an increased frequency of detachment after initial establishment of an end-on bound state. This indicates that KLP67A is important for maintaining stable kinetochore-microtubule attachments and proper chromosome alignment during mitosis. In order to test whether detachment frequency was enhanced due to the loss of KLP67A’s microtubule length regulation, S2 cells were treated with colcemid to destabilise microtubules in the absence of KLP67A. However, artificial shortening of the spindle was unable to rescue the phenotype, suggesting that KLP67A’s stabilising function at the kinetochore-microtubule interface was independent of its depolymerase activity. As KLP67A was also able to stabilise microtubule plus ends after inducing catastrophe in vitro, the authors hypothesised that this function may be important for the stabilisation of end-on attachments and thus could be potentially rescued with another plus-end stabilising factor. Ectopic expression of GFP-Mast/Orbit (CLASP in humans) in the absence of KLP67A was able suppress microtubule detachment from the kinetochore, but had no impact on the other effects of KLP67A depletion e.g. spindle elongation and mitotic duration.
Finally, the authors carried out their own RNAi analysis of Kif18a depletion in GFP-Mad2 expressing HeLa cell lines. GFP-Mad2 recruitment to the kinetochore was used as a read-out for incorrect attachments. Although no clear differences were observed between the localisation of Mad2 to unaligned kinetochores, recruitment of GFP-Mad2 to aligned kinetochores was doubled in Kif18a depleted cells. This suggests that Kif18a is required to maintain stable kinetochore-microtubule attachments similarly to KLP67A in Drosophila.
All in all, this preprint highlights the importance of KLP67A in (i) regulating microtubule dynamics within the spindle, (ii) stabilising the kinetochore-microtubule attachment and (iii) highlighting functional similarity to the roles of human Kif18a during mitosis.
Open Questions:
What effect does the KLP67A tail domain have on microtubule dynamics?
Can similar depolymerase and stabilisation activity be reconstituted for human Kif18a in vitro on dynamic microtubules to clarify the controversy around its function?
Are stabilising factors at the human kinetochore-microtubule interface, such as the Ska and SKAP/Astrin complex, responsible for dampening the severity of Kif18a depletion in human cells?
Further reading:
Goshima, G., and R.D. Vale. 2005. Cell cycle-dependent dynamics and regulation of mitotic kinesins in Drosophila S2 cells. Mol Biol Cell. 16:3896-3907.
Stumpff, J., Y. Du, C.A. English, Z. Maliga, M. Wagenbach, C.L. Asbury, L. Wordeman, and R. Ohi. 2011. A tethering mechanism controls the processivity and kinetochore-microtubule plus-end enrichment of the kinesin-8 Kif18A. Mol Cell. 43:764-775.
McHugh, T., A.A. Gluszek, and J.P.I. Welburn. 2018. Microtubule end tethering of a processive kinesin-8 motor Kif18b is required for spindle positioning. J Cell Biol. 217: 2403-2416
doi: https://doi.org/10.1242/prelights.5590
Read preprintSign up to customise the site to your preferences and to receive alerts
Register hereAlso in the biochemistry category:
Triglyceride metabolism controls inflammation and APOE4-associated disease states in microglia
Gustavo Stelzer, Marcus Oliveira
Impaired 26S proteasome causes learning and memory deficiency and induces neuroinflammation mediated by NF-κB in mice
Gustavo Stelzer, Marcus Oliveira
Notch3 is a genetic modifier of NODAL signalling for patterning asymmetry during mouse heart looping
Bhaval Parmar
Also in the cell biology category:
Platelet-derived LPA16:0 inhibits adult neurogenesis and stress resilience in anxiety disorder
Harvey Roweth
Investigating Mechanically Activated Currents from Trigeminal Neurons of Non-Human Primates
Vanessa Ehlers
Motor Clustering Enhances Kinesin-driven Vesicle Transport
Sharvari Pitke
preListsbiochemistry category:
in theBSCB-Biochemical Society 2024 Cell Migration meeting
This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.
List by | Reinier Prosee |
Peer Review in Biomedical Sciences
Communication of scientific knowledge has changed dramatically in recent decades and the public perception of scientific discoveries depends on the peer review process of articles published in scientific journals. Preprints are key vehicles for the dissemination of scientific discoveries, but they are still not properly recognized by the scientific community since peer review is very limited. On the other hand, peer review is very heterogeneous and a fundamental aspect to improve it is to train young scientists on how to think critically and how to evaluate scientific knowledge in a professional way. Thus, this course aims to: i) train students on how to perform peer review of scientific manuscripts in a professional manner; ii) develop students' critical thinking; iii) contribute to the appreciation of preprints as important vehicles for the dissemination of scientific knowledge without restrictions; iv) contribute to the development of students' curricula, as their opinions will be published and indexed on the preLights platform. The evaluations will be based on qualitative analyses of the oral presentations of preprints in the field of biomedical sciences deposited in the bioRxiv server, of the critical reports written by the students, as well as of the participation of the students during the preprints discussions.
List by | Marcus Oliveira et al. |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
List by | Nadja Hümpfer et al. |
20th “Genetics Workshops in Hungary”, Szeged (25th, September)
In this annual conference, Hungarian geneticists, biochemists and biotechnologists presented their works. Link: http://group.szbk.u-szeged.hu/minikonf/archive/prg2021.pdf
List by | Nándor Lipták |
Fibroblasts
The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!
List by | Osvaldo Contreras |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar et al. |
EMBL Seeing is Believing – Imaging the Molecular Processes of Life
Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019
List by | Dey Lab |
Cellular metabolism
A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.
List by | Pablo Ranea Robles |
MitoList
This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.
List by | Sandra Franco Iborra |
Also in the cell biology category:
November in preprints – the CellBio edition
This is the first community-driven preList! A group of preLighters, with expertise in different areas of cell biology, have worked together to create this preprint reading lists for researchers with an interest in cell biology. Categories include: 1) cancer cell biology 2) cell cycle and division 3) cell migration and cytoskeleton 4) cell organelles and organisation 5) cell signalling and mechanosensing 6) genetics/gene expression
List by | Felipe Del Valle Batalla et al. |
BSCB-Biochemical Society 2024 Cell Migration meeting
This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.
List by | Reinier Prosee |
‘In preprints’ from Development 2022-2023
A list of the preprints featured in Development's 'In preprints' articles between 2022-2023
List by | Alex Eve, Katherine Brown |
preLights peer support – preprints of interest
This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.
List by | preLights peer support |
The Society for Developmental Biology 82nd Annual Meeting
This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.
List by | Joyce Yu, Katherine Brown |
CSHL 87th Symposium: Stem Cells
Preprints mentioned by speakers at the #CSHLsymp23
List by | Alex Eve |
Journal of Cell Science meeting ‘Imaging Cell Dynamics’
This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.
List by | Helen Zenner |
9th International Symposium on the Biology of Vertebrate Sex Determination
This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.
List by | Martin Estermann |
Alumni picks – preLights 5th Birthday
This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.
List by | Sergio Menchero et al. |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
List by | Nadja Hümpfer et al. |
Fibroblasts
The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!
List by | Osvaldo Contreras |
EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)
A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.
List by | Alex Eve |
FENS 2020
A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020
List by | Ana Dorrego-Rivas |
Planar Cell Polarity – PCP
This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.
List by | Ana Dorrego-Rivas |
BioMalPar XVI: Biology and Pathology of the Malaria Parasite
[under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria
List by | Dey Lab, Samantha Seah |
1
Cell Polarity
Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.
List by | Yamini Ravichandran |
TAGC 2020
Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20
List by | Maiko Kitaoka et al. |
3D Gastruloids
A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.
List by | Paul Gerald L. Sanchez and Stefano Vianello |
ECFG15 – Fungal biology
Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome
List by | Hiral Shah |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar et al. |
EMBL Seeing is Believing – Imaging the Molecular Processes of Life
Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019
List by | Dey Lab |
Autophagy
Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.
List by | Sandra Malmgren Hill |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
List by | Rob Hynds |
Cellular metabolism
A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.
List by | Pablo Ranea Robles |
BSCB/BSDB Annual Meeting 2019
Preprints presented at the BSCB/BSDB Annual Meeting 2019
List by | Dey Lab |
Biophysical Society Annual Meeting 2019
Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA
List by | Joseph Jose Thottacherry |
ASCB/EMBO Annual Meeting 2018
This list relates to preprints that were discussed at the recent ASCB conference.
List by | Dey Lab, Amanda Haage |