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Structure of the fission yeast actomyosin ring during constriction

Swulius, Matthew T. and Nguyen, Lam T. and Ladinsky, Mark S. and Ortega, Davi R. and Aich, Samya and Mishra, Mithilesh and Jensen, Grant J. (2018) Structure of the fission yeast actomyosin ring during constriction. Proceedings of the National Academy of Sciences of the United States of America, 115 (7). E1455-E1464. ISSN 0027-8424. PMCID PMC5816141. https://resolver.caltech.edu/CaltechAUTHORS:20180119-110411055

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Abstract

Cell division in many eukaryotes is driven by a ring containing actin and myosin. While much is known about the main proteins involved, the precise arrangement of actin filaments within the contractile machinery, and how force is transmitted to the membrane, remains unclear. Here we use cryosectioning and cryofocused ion beam milling to gain access to cryopreserved actomyosin rings in Schizosaccharomyces pombe for direct 3D imaging by electron cryotomography. Our results show that straight, overlapping actin filaments, running nearly parallel to each other and to the membrane, form a loose bundle of ∼150 nm in diameter that “saddles” the inward-bending membrane at the leading edge of the division septum. The filaments do not make direct contact with the membrane. Our analysis of the actin filaments reveals the variability in filament number, nearest-neighbor distances between filaments within the bundle, their distance from the membrane, and angular distribution with respect to the membrane.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1073/pnas.1711218115DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5816141/PubMed CentralArticle
http://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1711218115/-/DCSupplementalPublisherSupporting Information
https://doi.org/10.1101/194902DOIDiscussion Paper
ORCID:
AuthorORCID
Ladinsky, Mark S.0000-0002-1036-3513
Ortega, Davi R.0000-0002-8344-2335
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2018 National Academy of Sciences. Published under the PNAS license. Edited by Thomas D. Pollard, Yale University, New Haven, CT, and approved December 13, 2017 (received for review June 23, 2017). Published online before print January 18, 2018. We thank Catherine Oikonomou for revising the manuscript for clarity and Frances Allen for initial training and guidance in cryo-FIB milling. We would also like to thank the reviewers for all of their critical feedback during the refinement of the manuscript. M.M. is an Intermediate Fellow of the Wellcome Trust−Department of Biotechnology India Alliance (IA/I/14/1/501317). M.M. acknowledges the India Alliance and the Department of Atomic Energy/Tata Institute of Fundamental Research for funds. This work was supported in part by National Institutes of Heath Grants GM122588 and GM082545 (to G.J.J.). Author contributions: M.T.S., L.T.N., M.M., and G.J.J. designed research; M.T.S., L.T.N., and M.S.L. performed research; D.R.O. and S.A. contributed new reagents/analytic tools; M.T.S., L.T.N., M.M., and G.J.J. analyzed data; and M.T.S., L.T.N., M.M., and G.J.J. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1711218115/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
Wellcome Trust−Department of Biotechnology India AllianceIA/I/14/1/501317
India AllianceUNSPECIFIED
Department of Atomic Energy (India)UNSPECIFIED
Tata Institute of Fundamental ResearchUNSPECIFIED
NIHGM122588
NIHGM082545
Subject Keywords:cell division; cytokinesis; actomyosin ring; cryoEM; FIB mill
Issue or Number:7
PubMed Central ID:PMC5816141
Record Number:CaltechAUTHORS:20180119-110411055
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180119-110411055
Official Citation:Structure of the fission yeast actomyosin ring, Matthew T. Swulius, Lam T. Nguyen, Mark S. Ladinsky, Davi R. Ortega, Samya Aich, Mithilesh Mishra, Grant J. Jensen. Proceedings of the National Academy of Sciences Feb 2018, 115 (7) E1455-E1464; DOI: 10.1073/pnas.1711218115.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84419
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:19 Jan 2018 21:43
Last Modified:05 Nov 2019 20:50

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