scholarly journals Live-cell single-molecule tracking reveals co-recognition of H3K27me3 and DNA targets polycomb Cbx7-PRC1 to chromatin

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Chao Yu Zhen ◽  
Roubina Tatavosian ◽  
Thao Ngoc Huynh ◽  
Huy Nguyen Duc ◽  
Raibatak Das ◽  
...  
Keyword(s):  
Single Molecule ◽  
Molecular Mechanisms ◽  
Histone H3 ◽  
Live Cell ◽  
Disease Pathogenesis ◽  
Single Molecule Tracking ◽  
Cooperation Mechanism ◽  
Functional Dna ◽  
Biochemical Analyses ◽  
Prevailing View

The Polycomb PRC1 plays essential roles in development and disease pathogenesis. Targeting of PRC1 to chromatin is thought to be mediated by the Cbx family proteins (Cbx2/4/6/7/8) binding to histone H3 with a K27me3 modification (H3K27me3). Despite this prevailing view, the molecular mechanisms of targeting remain poorly understood. Here, by combining live-cell single-molecule tracking (SMT) and genetic engineering, we reveal that H3K27me3 contributes significantly to the targeting of Cbx7 and Cbx8 to chromatin, but less to Cbx2, Cbx4, and Cbx6. Genetic disruption of the complex formation of PRC1 facilitates the targeting of Cbx7 to chromatin. Biochemical analyses uncover that the CD and AT-hook-like (ATL) motif of Cbx7 constitute a functional DNA-binding unit. Live-cell SMT of Cbx7 mutants demonstrates that Cbx7 is targeted to chromatin by co-recognizing of H3K27me3 and DNA. Our data suggest a novel hierarchical cooperation mechanism by which histone modifications and DNA coordinate to target chromatin regulatory complexes.

scholarly journals Structural Contributions to Hydrodynamic Diameter for Quantum Dots Optimized for Live-Cell Single-Molecule Tracking

2018 ◽  
Vol 122 (30) ◽  
pp. 17406-17412 ◽  
Author(s):  
Janet Y. Sheung ◽  
Pinghua Ge ◽  
Sung Jun Lim ◽  
Sang Hak Lee ◽  
Andrew M. Smith ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Single Molecule ◽  
Live Cell ◽  
Hydrodynamic Diameter ◽  
Single Molecule Tracking

scholarly journals Single-molecule imaging of Hedgehog pathway protein Smoothened in primary cilia reveals binding events regulated by Patched1

2015 ◽  
Vol 112 (27) ◽  
pp. 8320-8325 ◽  
Author(s):  
Ljiljana Milenkovic ◽  
Lucien E. Weiss ◽  
Joshua Yoon ◽  
Theodore L. Roth ◽  
YouRong S. Su ◽  
...  
Keyword(s):  
Single Molecule ◽  
Binding Sites ◽  
Primary Cilia ◽  
Molecular Mechanisms ◽  
Single Molecule Tracking ◽  
Temporal Precision ◽  
Essential Step ◽  
Pathway Activation ◽  
Patched 1 ◽  
Diffusive Movement

Accumulation of the signaling protein Smoothened (Smo) in the membrane of primary cilia is an essential step in Hedgehog (Hh) signal transduction, yet the molecular mechanisms of Smo movement and localization are poorly understood. Using ultrasensitive single-molecule tracking with high spatial/temporal precision (30 nm/10 ms), we discovered that binding events disrupt the primarily diffusive movement of Smo in cilia at an array of sites near the base. The affinity of Smo for these binding sites was modulated by the Hh pathway activation state. Activation, by either a ligand or genetic loss of the negatively acting Hh receptor Patched-1 (Ptch), reduced the affinity and frequency of Smo binding at the base. Our findings quantify activation-dependent changes in Smo dynamics in cilia and highlight a previously unknown step in Hh pathway activation.

scholarly journals Correction: Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins

Nanoscale ◽  
2020 ◽  
Vol 12 (35) ◽  
pp. 18476-18477
Author(s):  
Andres I. König ◽  
Raya Sorkin ◽  
Ariel Alon ◽  
Dikla Nachmias ◽  
Kalyan Dhara ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Single Molecule ◽  
Live Cell ◽  
Plasma Membrane Proteins ◽  
Single Molecule Tracking ◽  
Localization Microscopy

Correction for ‘Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins’ by Andres I. König et al., Nanoscale, 2020, 12, 3236–3248, DOI: 10.1039/C9NR08594G.

Deep-Learning-Assisted Single-Molecule Tracking on a Live Cell Membrane

2021 ◽  
Author(s):  
Qian Wang ◽  
Hua He ◽  
Qian Zhang ◽  
Zhenzhen Feng ◽  
Jiqiang Li ◽  
...  
Keyword(s):  
Deep Learning ◽  
Cell Membrane ◽  
Single Molecule ◽  
Live Cell ◽  
Single Molecule Tracking
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