| Title: |
RNA binding proteins Smaug and Cup induce CCR4–NOT-dependent deadenylation of the nanos mRNA in a reconstituted system |
| Authors: |
Pekovic, Filip; Rammelt, Christiane; Kubíková, Jana; Metz, Jutta; Jeske, Mandy; Wahle, Elmar |
| Publication Year: |
2023 |
| Collection: |
Share it - Open Access und Forschungsdaten-Repositorium der Hochschulbibliotheken in Sachsen-Anhalt |
| Subject Terms: |
ddc:570; ddc:572 |
| Description: |
Posttranscriptional regulation of the maternal nanos mRNA is essential for the development of the anterior – posterior axis of the Drosophila embryo. The nanos RNA is regulated by the protein Smaug, which binds to Smaug recognition elements (SREs) in the nanos 3’-UTR and nucleates the assembly of a larger repressor complex including the eIF4E-T paralog Cup and five additional proteins. The Smaug-dependent complex represses translation of nanos and induces its deadenylation by the CCR4–NOT deadenylase. Here we report an in vitro reconstitution of the Drosophila CCR4–NOT complex and Smaug-dependent deadenylation. We find that Smaug by itself is sufficient to cause deadenylation by the Drosophila or human CCR4–NOT complexes in an SRE-dependent manner. CCR4–NOT subunits NOT10 and NOT11 are dispensable, but the NOT module, consisting of NOT2, NOT3 and the C-terminal part of NOT1, is required. Smaug interacts with the C-terminal domain of NOT3. Both catalytic subunits of CCR4–NOT contribute to Smaug-dependent deadenylation. Whereas the CCR4–NOT complex itself acts distributively, Smaug induces a processive behavior. The cytoplasmic poly(A) binding protein (PABPC) has a minor inhibitory effect on Smaug-dependent deadenylation. Among the additional constituents of the Smaug-dependent repressor complex, Cup also facilitates CCR4–NOT-dependent deadenylation, both independently and in cooperation with Smaug. |
| Document Type: |
article in journal/newspaper |
| File Description: |
application/pdf |
| Language: |
English |
| ISBN: |
978-1-84537-774-8; 1-84537-774-5 |
| Relation: |
http://dx.doi.org/10.25673/103303 |
| DOI: |
10.25673/103303 |
| Availability: |
https://opendata.uni-halle.de//handle/1981185920/105255; https://doi.org/10.25673/103303 |
| Rights: |
https://creativecommons.org/licenses/by/4.0/ |
| Accession Number: |
edsbas.2E1F782F |
| Database: |
BASE |