Mouse dead end 1-β interacts with c-Jun and stimulates activator protein 1 transactivation
Yong Zhang1, Yan-Lin Su1, Le-Sai Li2
1Department of Internal Medicine, College of Medicine, Hunan Normal University, Changsha, Hunan 410013, P.R. China.
Abstract:
Dead end 1 (DND1), important for maintaining the viability of primordial germ cells, is the first protein containing an RNA recognition motif that has been directly implicated as a heritable cause of spontaneous tumorigenesis. In the present study, c-Jun was identified through yeast two-hybrid screening of a 10.5-day old mouse embryo cDNA library as one of the proteins which interact with DND1-β. The interaction between DND1-β and c-Jun was demonstrated to occur by glutathione S‑transferase pull‑down and co-immunoprecipitation. Using confocal microscopy, DND1-β was found to be specifically expressed in GC-1 spermatogonia cells, mainly in the nuclei. When transfected into GC-1 cells, DND1-β and c-Jun were demonstrated to be co-localized principally in the nuclei. Furthermore, in a dual luciferase reporter assay, the transcriptional activity of activator protein 1 was demonstrated to be significantly increased by co-transfection with DND1-β and c-Jun plasmids in GC-1 cells. The identification and confirmation of an additional protein interacting with DND1-β facilitates the investigation of the functions and molecular mechanisms of DND1.
Insights
Dead end 1 (DND1), a protein linked to tumorigenesis, interacts with c-Jun. This interaction, confirmed in mouse spermatogonia cells, enhances transcriptional activity, aiding DND1 function studies.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Dead end 1 (DND1) is crucial for primordial germ cell viability and is implicated in heritable tumorigenesis.
- DND1 is the first identified protein with an RNA recognition motif linked to spontaneous cancer.
- Understanding DND1's molecular interactions is key to elucidating its role in tumorigenesis.
Purpose of the Study:
- To identify proteins interacting with DND1-β.
- To investigate the functional consequences of DND1-β and c-Jun interaction.
- To explore the role of DND1 in gene transcription.
Main Methods:
- Yeast two-hybrid screening using a mouse embryo cDNA library.
- Glutathione S-transferase pull-down and co-immunoprecipitation assays.
- Confocal microscopy for protein localization.
- Dual luciferase reporter assay for transcriptional activity.
Main Results:
- c-Jun was identified as a DND1-β interacting protein.
- DND1-β and c-Jun co-localize in the nuclei of GC-1 spermatogonia cells.
- Co-transfection of DND1-β and c-Jun significantly increased activator protein 1 transcriptional activity.
Conclusions:
- This study identifies c-Jun as a novel interacting partner of DND1-β.
- The interaction between DND1-β and c-Jun influences transcriptional regulation.
- These findings provide new insights into the molecular mechanisms of DND1 function in cellular processes.
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