Interactions of drosophila cryptochrome.

APEX2 TurboID cryptochrome photoreceptor protein-protein interactions proteomics

Journal

Photochemistry and photobiology
ISSN: 1751-1097
Titre abrégé: Photochem Photobiol
Pays: United States
ID NLM: 0376425

Informations de publication

Date de publication:
05 Feb 2024
Historique:
revised: 20 01 2024
received: 14 09 2023
accepted: 23 01 2024
medline: 5 2 2024
pubmed: 5 2 2024
entrez: 5 2 2024
Statut: aheadofprint

Résumé

In this study, we investigate the intricate regulatory mechanisms underlying the circadian clock in Drosophila, 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 spectrometry. 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, our 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 investigations into light- and time-dependent protein interactions in Drosophila.

Identifiants

pubmed: 38314442
doi: 10.1111/php.13916
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : The Scientific and Technological Research Council of Turkiye (TUBITAK)
ID : 219Z101

Informations de copyright

© 2024 American Society for Photobiology.

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Auteurs

Gozde Ozcelik (G)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Mehmet Serdar Koca (MS)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Buket Sunbul (B)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Fatma Yilmaz-Atay (F)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Feride Demirhan (F)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Busra Tiryaki (B)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Kevser Cilenk (K)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Saba Selvi (S)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Nuri Ozturk (N)

Department of Molecular Biology and Genetics, Gebze Technical University, Gebze, Kocaeli, Turkey.

Classifications MeSH