Motional dynamics of single Patched1 molecules in cilia are controlled by Hedgehog and cholesterol

Proceedings of the National Academy of Sciences of the United States of America
Lucien E WeissW E Moerner

Abstract

The Hedgehog-signaling pathway is an important target in cancer research and regenerative medicine; yet, on the cellular level, many steps are still poorly understood. Extensive studies of the bulk behavior of the key proteins in the pathway established that during signal transduction they dynamically localize in primary cilia, antenna-like solitary organelles present on most cells. The secreted Hedgehog ligand Sonic Hedgehog (SHH) binds to its receptor Patched1 (PTCH1) in primary cilia, causing its inactivation and delocalization from cilia. At the same time, the transmembrane protein Smoothened (SMO) is released of its inhibition by PTCH1 and accumulates in cilia. We used advanced, single molecule-based microscopy to investigate these processes in live cells. As previously observed for SMO, PTCH1 molecules in cilia predominantly move by diffusion and less frequently by directional transport, and spend a fraction of time confined. After treatment with SHH we observed two major changes in the motional dynamics of PTCH1 in cilia. First, PTCH1 molecules spend more time as confined, and less time freely diffusing. This result could be mimicked by a depletion of cholesterol from cells. Second, after treatment with SHH, but not afte...Continue Reading

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Citations

Oct 30, 2019·Proceedings of the National Academy of Sciences of the United States of America·Nicholas A MoringoChristy F Landes
Dec 8, 2020·The Journal of Cell Biology·Sarah FindaklyDavid R Raleigh
Nov 18, 2020·Nature Chemical Biology·Arun RadhakrishnanChristian Siebold
Oct 10, 2020·Journal of the American Chemical Society·Leonhard Möckl, W E Moerner
May 4, 2021·Journal of Extracellular Vesicles·Ashraf M MohieldinSurya M Nauli
Jun 11, 2021·Biochemical Pharmacology·Vikas DaggubatiNavdar Sever
Aug 14, 2021·Nature Communications·Carla E Cadena Del CastilloAnne Spang
Aug 28, 2021·International Journal of Molecular Sciences·Laurent FernandezPierre-Emmanuel Milhiet
Sep 16, 2021·Journal of Chemical Theory and Computation·T Bertie AnsellRobin A Corey
Nov 5, 2021·Proceedings of the National Academy of Sciences of the United States of America·Zhiqing ZhangErwin J G Peterman
Jan 11, 2022·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Tara AkhshiWilliam S Trimble

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Methods Mentioned

BETA
confocal microscopy

Software Mentioned

MATLAB scripts
lsqnonlin

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