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Method Article
The use of Optical Magnetic Twisting Cytometry and Flourescence Resonance Energy Transfer to quantify force-induced protein dissociation in the nucleus of a living cell
Ning Wang
University of Illnois
Corresponding Author
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Mechanical forces are known to play a significant role in biological processes. These forces can be transmitted to the cell through the cytoskeletal filament network, inducing different biochemical responses within the cytoplasm. Although there have been ample reports showing that cytoplasmic enzymes can be directly activated by a local stress on the cell surface via integrins, there has been no evidence that mechanical forces can directly alter nuclear functions without intermediate biochemical cascades. Recently, we showed evidence that forces on the cell membrane can be transmitted directly into the nucleus, inducing structural changes of protein complexes in Cajal bodies. Here, we describe a protocol that utilizes the optical magnetic twisting cytometry (MTC) for force application and fluorescent resonance energy transfer (FRET) to monitor the dynamics and interaction of various Cajal body proteins.
Keywords
Mechanical forces, nuclear function, nuclear organization, Cajal body, mechanotransduction
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Associated Publications
Dynamic force-induced direct dissociation of protein complexes in a nuclear body in living cells
by Yeh-Chuin Poh, Sergey P. Shevtsov, Farhan Chowdhury, +4
Nature Communications (09 April, 2012)
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This is a preprint. It has not completed peer review.
Method Article
The use of Optical Magnetic Twisting Cytometry and Flourescence Resonance Energy Transfer to quantify force-induced protein dissociation in the nucleus of a living cell
Ning Wang
University of Illnois
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 06 Jun, 2012
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Mechanical forces are known to play a significant role in biological processes. These forces can be transmitted to the cell through the cytoskeletal filament network, inducing different biochemical responses within the cytoplasm. Although there have been ample reports showing that cytoplasmic enzymes can be directly activated by a local stress on the cell surface via integrins, there has been no evidence that mechanical forces can directly alter nuclear functions without intermediate biochemical cascades. Recently, we showed evidence that forces on the cell membrane can be transmitted directly into the nucleus, inducing structural changes of protein complexes in Cajal bodies. Here, we describe a protocol that utilizes the optical magnetic twisting cytometry (MTC) for force application and fluorescent resonance energy transfer (FRET) to monitor the dynamics and interaction of various Cajal body proteins.
Figure 1
Associated Publications
Dynamic force-induced direct dissociation of protein complexes in a nuclear body in living cells
by Yeh-Chuin Poh, Sergey P. Shevtsov, Farhan Chowdhury, +4
Nature Communications (09 April, 2012)