bei Google ScholarTitelangaben
Loedige, Inga ; Jakob, Leonhard ; Treiber, Thomas ; Ray, Debashish ; Stotz, Mathias ; Treiber, Nora ; Hennig, Janosch ; Cook, Kate B. ; Morris, Quaid ; Hughes, Timothy R. ; Engelmann, Julia C. ; Krahn, Michael P. ; Meister, Gunter:
The Crystal Structure of the NHL Domain in Complex with RNA Reveals the Molecular Basis of Drosophila Brain-Tumor-Mediated Gene Regulation.
In: Cell Reports.
Bd. 13
(2015)
Heft 6
.
- S. 1206-1220.
ISSN 2211-1247
DOI: https://doi.org/10.1016/j.celrep.2015.09.068
Abstract
TRIM-NHL proteins are conserved among metazoans and control cell fate decisions in various stem cell linages. The Drosophila TRIM-NHL protein Brain tumor (Brat) directs differentiation of neuronal stem cells by suppressing self-renewal factors. Brat is an RNA-binding protein and functions as a translational repressor. However, it is unknown which RNAs Brat regulates and how RNA-binding specificity is achieved. Using RNA immunoprecipitation and RNAcompete, we identify Brat-bound mRNAs in Drosophila embryos and define consensus binding motifs for Brat as well as a number of additional TRIM-NHL proteins, indicating that TRIM-NHL proteins are conserved, sequence-specific RNA-binding proteins. We demonstrate that Brat-mediated repression and direct RNA-binding depend on the identified motif and show that binding of the localization factor Miranda to the Brat-NHL domain inhibits Brat activity. Finally, to unravel the sequence specificity of the NHL domain, we crystallize the Brat-NHL domain in complex with RNA and present a high-resolution protein-RNA structure of this fold.