TY  - INPR
A1  - Keil, Philipp
A1  - Wulf, Alexander
A1  - Kachariya, Nitin
A1  - Reuscher, Samira
A1  - Hühn, Kristin
A1  - Silbern, Ivan
A1  - Altmüller, Janine
A1  - Stehle, Ralf
A1  - Zarnack, Katharina
A1  - Sattler, Michael
A1  - Urlaub, Henning
A1  - Sträßer, Katja
T1  - Npl3 functions in mRNP assembly by recruitment of mRNP components to the transcription site and their transfer onto the mRNA
T2  - bioRxiv
N2  - RNA-binding proteins (RBPs) control every RNA metabolic process by multiple protein-RNA and protein-protein interactions. Their roles have largely been analyzed by crude mutations, which abrogate multiple functions at once and likely impact the structural integrity of the large messenger ribonucleoprotein particle (mRNP) assemblies, these proteins often function in. Using UV-induced RNA-protein crosslinking and subsequent mass spectrometric analysis, we first identified more than 100 in vivo RNA crosslinks in 16 nuclear mRNP components in S. cerevisiae. For functional analysis, we chose Npl3, for which we determined crosslinks in its two RNA recognition motifs (RRM) and in the flexible linker region connecting the two. Using NMR and structural analyses, we show that both RRM domains and the linker uniquely contribute to RNA recognition. Interestingly, mutations in these regions cause different phenotypes, indicating distinct functions of the different RNA-binding domains of Npl3. Notably, the npl3-Linker mutation strongly impairs recruitment of several mRNP components to chromatin and incorporation of further mRNP components into nuclear mRNPs, establishing a function of Npl3 in nuclear mRNP assembly. Taken together, we determined the specific function of the RNA-binding activity of the nuclear mRNP component Npl3, an approach that can be applied to many RBPs in any RNA metabolic process.
Y1  - 2022
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/73083
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-730837
UR  - https://www.biorxiv.org/content/10.1101/2022.07.22.501171v1
IS  - 2022.07.22.501171 Version 1
PB  - bioRxiv
ER  -