Reversible Optical Control of Receptor Tyrosine Kinase Activity and ERK Dynamics Using Azobenzene-Carrying DNA...

Area of Science:

• Molecular Biology
• Cell Signaling
• Biotechnology

Background:

• Receptor tyrosine kinases (RTKs) are crucial for cell signaling and cell fate determination.
• Temporal dynamics of RTK activity significantly influence cellular responses.
• Existing methods for precise, non-genetic control of endogenous RTKs are limited.

Purpose of the Study:

• To develop a method for high-temporal-resolution control of endogenous RTK activity.
• To investigate the role of RTK activation dynamics in cell signaling and fate.
• To create a tool for precise modulation of c-Met receptor activity.

Main Methods:

• Designed a DNA aptamer agonist (Met-azo-aptamer) incorporating a photoisomerizable azobenzene molecule.
• Engineered the aptamer for reversible optical control of c-Met receptor binding and activity.
• Applied the Met-azo-aptamer to HeLa cells and used UV/blue light to modulate c-Met activity.

Main Results:

• The Met-azo-aptamer enabled light-induced activation and deactivation of c-Met receptor signaling.
• Cellular phosphorylation signals were modulated in response to specific light patterns.
• The timing, amplitude, and duration of downstream ERK activation were precisely controlled by varying light patterns.

Conclusions:

• Met-azo-aptamer provides reversible optical control over c-Met receptor activity.
• This tool offers a high-resolution method to study RTK activation dynamics.
• The findings facilitate research into the relationship between RTK signaling patterns and cell function/fate.