scholarly journals Quantitative analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy

2019 ◽  
Author(s):  
Matthias Altmeyer ◽  
Jone Michelena ◽  
Aleksandra Lezaja ◽  
Federico Teloni ◽  
Thomas Schmid ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Fluorescence Microscopy ◽  
Parp Inhibitor ◽  
Multidimensional Fluorescence

scholarly journals Analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy reveals mechanisms of sensitivity and resistance

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Jone Michelena ◽  
Aleksandra Lezaja ◽  
Federico Teloni ◽  
Thomas Schmid ◽  
Ralph Imhof ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Parp Inhibitor ◽  
Multidimensional Fluorescence

scholarly journals Divergent modes for cargo-mediated control of clathrin-coated pit dynamics

2013 ◽  
Vol 24 (11) ◽  
pp. 1725-1734 ◽  
Author(s):  
Amanda L. Soohoo ◽  
Manojkumar A. Puthenveedu
Keyword(s):  
Quantitative Analysis ◽  
Fluorescence Microscopy ◽  
Total Internal Reflection ◽  
Live Cells ◽  
Coated Pits ◽  
Microscopy Imaging ◽  
Cargo Proteins ◽  
Μ Opioid Receptor ◽  
Signaling Receptors ◽  
G Protein Coupled

Clathrin-mediated endocytosis has long been viewed as a process driven by core endocytic proteins, with internalized cargo proteins being passive. In contrast, an emerging view suggests that signaling receptor cargo may actively control its fate by regulating the dynamics of clathrin-coated pits (CCPs) that mediate their internalization. Despite its physiological implications, very little is known about such “cargo-mediated regulation” of CCPs by signaling receptors. Here, using multicolor total internal reflection fluorescence microscopy imaging and quantitative analysis in live cells, we show that the μ-opioid receptor, a physiologically relevant G protein–coupled signaling receptor, delays the dynamics of CCPs in which it is localized. This delay is mediated by the interactions of two critical leucines on the receptor cytoplasmic tail. Unlike the previously known mechanism of cargo-mediated regulation, these residues regulate the lifetimes of dynamin, a key component of CCP scission. These results identify a novel means for selectively controlling the endocytosis of distinct cargo that share common trafficking components and indicate that CCP regulation by signaling receptors can operate via divergent modes.

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