Interactions of drosophila cryptochrome

In this study, we investigate the intricate regulatory mechanisms underlying the circadian clock inDrosophila, focusing on the light-induced conformational changes in the cryptochrome (DmCry). Upon light exposure, DmCry undergoes conformational changes that prompt its binding to Timeless and Jetlag proteins, initiating a cascade crucial for the starting of a new circadian cycle. DmCry is subsequently degraded, contributing to the desensitization of the resetting mechanism. The transient and short-lived nature of DmCry protein –protein interactions (PPIs), leading to DmCry degradation within an hour of light exposure, presents a challenge for comprehensive exploration. To address this, we employed proximity-dependent biotinylation techniques, combining engineered BioID (TurboID) and APEX (APEX2) enzymes with mass spectr ometry. This approach enabled the identification of the in vitro DmCry interactome in Drosophila S2 cells, uncovering several novel PPIs associated with DmCry. Validation of these interactions through a novel co-immunoprecipitation technique enhances the reliability of our findings. Importantly, ou r study suggests the potential of this method to reveal additional circadian clock- or magnetic field-dependent PPIs involving DmCry. This exploration of the DmCry interactome not only advances our understanding of circadian clock regulation but also establishes a versatile framework for future inve stigations into light- and time-dependent protein interactions in...

Source: Photochemistry and Photobiology - February 5, 2024 Category: Science Authors: Gozde Ozcelik, Mehmet Serdar Koca, Buket Sunbul, Fatma Yilmaz ‐Atay, Feride Demirhan, Busra Tiryaki, Kevser Cilenk, Saba Selvi, Nuri Ozturk Tags: RESEARCH ARTICLE Source Type: research