Optogenetic dissection of mitotic spindle positioning in vivo
Daniel J. Dickinson,
Anna A. Akhmanova,
Sander J.L. van den Heuvel
Posted 14 May 2018
bioRxiv DOI: 10.1101/319772 (published DOI: 10.7554/elife.38198)
Posted 14 May 2018
The position of the mitotic spindle determines the plane of cell cleavage, and thereby the location, size, and content of daughter cells. Spindle positioning is driven by dynein-mediated pulling forces exerted on astral microtubules. This process requires an evolutionarily conserved complex of α-GDP, GPR-1,2/Pins/LGN, and LIN-5/Mud/NuMA proteins. It remains unknown whether this complex merely forms a membrane anchor for dynein, or whether the individual components have additional functions, for instance through Gα-GTP or dynein activation. To functionally dissect this system, we developed a genetic strategy for light-controlled localization of endogenous proteins in C. elegans embryos. Controlled germline expression and membrane recruitment of the Gα; regulators RIC-8/Ric-8A and RGS-7/Loco/RGS3, and replacement of Gα with a light-inducible membrane anchor demonstrated that Gα-GTP signaling is dispensable for pulling force generation. In the absence of Gα, cortical recruitment of GPR-1,2 or LIN-5, but not dynein itself, induced high pulling forces. Local recruitment of LIN-5 overruled normal cell-cycle and polarity regulation, and provided experimental control over the spindle and cell cleavage plane. Our results define Gα-GDP-GPR-1,2/Pins/LGN as a regulatable membrane anchor, and LIN-5/Mud/NuMA as a potent activator of dynein-dependent spindle positioning forces. This study also highlights the possibilities for optogenetic control of endogenous proteins within an animal system.
- Downloaded 526 times
- Download rankings, all-time:
- Site-wide: 30,620 out of 92,758
- In cell biology: 1,450 out of 4,781
- Year to date:
- Site-wide: 74,808 out of 92,758
- Since beginning of last month:
- Site-wide: 65,521 out of 92,758
Downloads over time
Distribution of downloads per paper, site-wide
- 18 Dec 2019: We're pleased to announce PanLingua, a new tool that enables you to search for machine-translated bioRxiv preprints using more than 100 different languages.
- 21 May 2019: PLOS Biology has published a community page about Rxivist.org and its design.
- 10 May 2019: The paper analyzing the Rxivist dataset has been published at eLife.
- 1 Mar 2019: We now have summary statistics about bioRxiv downloads and submissions.
- 8 Feb 2019: Data from Altmetric is now available on the Rxivist details page for every preprint. Look for the "donut" under the download metrics.
- 30 Jan 2019: preLights has featured the Rxivist preprint and written about our findings.
- 22 Jan 2019: Nature just published an article about Rxivist and our data.
- 13 Jan 2019: The Rxivist preprint is live!