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  MINFLUX reveals dynein stepping in live neurons

Schleske, J. M., Hubrich, J., Wirth, J. O., D'Este, E., Engelhardt, J., & Hell, S. W. (2024). MINFLUX reveals dynein stepping in live neurons. Proceedings of the National Academy of Sciences of the United States of America, 121(38): e2412241121, pp. 1-8. doi:10.1073/pnas.2412241121.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000F-DF02-4 Version Permalink: https://hdl.handle.net/21.11116/0000-000F-E3B3-6
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 Creators:
Schleske, Jonas M.1, Author           
Hubrich, Jasmine1, Author           
Wirth, Jan Otto1, Author           
D'Este, Elisa1, Author           
Engelhardt, Johann1, Author           
Hell, Stefan W.1, Author                 
Affiliations:
1Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society, ou_2364730              

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Free keywords: CRISPR/Cas9; MINFLUX; dynein; fluorescence nanoscopy; live neurons
 Abstract: Dynein is the primary molecular motor responsible for retrograde intracellular transport of a variety of cargoes, performing successive nanometer-sized steps within milliseconds. Due to the limited spatiotemporal precision of established methods for molecular tracking, current knowledge of dynein stepping is essentially limited to slowed-down measurements in vitro. Here, we use MINFLUX fluorophore localization to directly track CRISPR/Cas9-tagged endogenous dynein with nanometer/millisecond precision in living primary neurons. We show that endogenous dynein primarily takes 8 nm steps, including frequent sideways steps but few backward steps. Strikingly, the majority of direction reversals between retrograde and anterograde movement occurred on the time scale of single steps (16 ms), suggesting a rapid regulatory reversal mechanism. Tug-of-war-like behavior during pauses or reversals was unexpectedly rare. By analyzing the dwell time between steps, we concluded that a single rate-limiting process underlies the dynein stepping mechanism, likely arising from just one adenosine 5'-triphosphate hydrolysis event being required during each step. Our study underscores the power of MINFLUX localization to elucidate the spatiotemporal changes underlying protein function in living cells.

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Language(s): eng - English
 Dates: 2024-06-182024-08-132024-09-10
 Publication Status: Published online
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: URI: https://pubmed.ncbi.nlm.nih.gov/39254993/
DOI: 10.1073/pnas.2412241121
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Title: Proceedings of the National Academy of Sciences of the United States of America
  Other : PNAS
  Other : Proceedings of the National Academy of Sciences of the USA
  Abbreviation : Proc. Natl. Acad. Sci. U. S. A.
Source Genre: Journal
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Publ. Info: Washington, D.C. : National Academy of Sciences
Pages: - Volume / Issue: 121 (38) Sequence Number: e2412241121 Start / End Page: 1 - 8 Identifier: ISSN: 0027-8424
CoNE: https://pure.mpg.de/cone/journals/resource/954925427230