Figure data for PNAS publication: Motional dynamics of single Patched1 molecules in cilia are controlled by Hedgehog and cholesterol by Lucien E. Weiss, Ljiljana Milenkovic, Josh Yoon, Tim Stearns, W. E. Moerner
- Figures for this research: 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 after cholesterol depletion, the molecules that remain in the diffusive state showed a significant increase in the diffusion coefficient. Therefore, PTCH1 inactivation by SHH changes the diffusive motion of PTCH1, possibly by modifying the membrane microenvironment in which PTCH1 resides.
|Type of resource
|[ca. 2017 - 2019]
|Weiss, Lucien E
|Ljiljana Milenkovic, Lucien E. Weiss, Joshua Yoon, Theodore L. Roth, YouRong S. Su, Steffen J. Sahl, Matthew P. Scott, and W. E. Moerner, “Single-molecule imaging of Hedgehog pathway protein Smoothened in primary cilia reveals binding events regulated by Patched1,” Proc. Nat. Acad. Sci. (USA) 112, 8320-8325 (2015) (https://doi.org/10.1073/pnas.1510094112, published online June 22, 2015).
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- Preferred Citation
Weiss, Lucien E., Milenkovic, Ljiljana, Yoon, Joshua, Stearns, Tim, and Moerner, W. E. (2019). Figure data for PNAS publication: Motional dynamics of single Patched1 molecules in cilia are controlled by Hedgehog and cholesterol
by Lucien E. Weiss, Ljiljana Milenkovic, Josh Yoon, Tim Stearns, W. E. Moerner. Stanford Digital Repository. Available at: https://purl.stanford.edu/sh175by4999
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