PXD035153

PXD035153 is an original dataset announced via ProteomeXchange.

Title	Npl3 functions in mRNP assembly by recruitment of mRNP components to the transcription site and their transfer onto the mRNA
Description	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 several functions at once and likely impact the structural integrity of the large mRNP assemblies, these proteins often function in. Thus, the function of the RNA-binding activity of RBPs is often unknown. Using UV-induced RNA-protein crosslinking and subsequent mass spectrometric (MS) analysis, we first identified more than 100 in vivo RNA crosslinks in 16 nuclear mRNP components. For functional analysis, we chose Npl3, for which we identified crosslinks in its two RNA recognition motifs (RRMs) as well as in the flexible linker connecting the two. Both RRM domains and the linker uniquely contribute to RNA recognition. Interestingly, mutations in each of these three regions cause different phenotypes, indicating distinct functions of the different RNA-binding domains of Npl3. Notably, the npl3-Linker mutant strongly impairs recruitment of some mRNP components to chromatin and the incorporation of other mRNP components into nuclear mRNPs, establishing a function of Npl3 in mRNP assembly. Taken together, we determined the specific function of the RNA-binding activity of a nuclear mRNP component, an approach that can be applied to many RBPs in any RNA metabolic process.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:36:51.817.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Alexander Wulf
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-07 01:08:15	ID requested
1	2023-03-10 20:20:27	announced
⏵ 2	2023-11-14 08:36:54	announced	2023-11-14: Updated project metadata.

Keil P, Wulf A, Kachariya N, Reuscher S, H, ü, hn K, Silbern I, Altm, ü, ller J, Keller M, Stehle R, Zarnack K, Sattler M, Urlaub H, Str, ä, ß, er K, Npl3 functions in mRNP assembly by recruitment of mRNP components to the transcription site and their transfer onto the mRNA. Nucleic Acids Res, 51(2):831-851(2023) [pubmed]

submitter keyword: RNA-binding sites, Npl3, SR-like protein, mRNP assembly, structure, XL-MS, RNA-binding proteins, NMR, mRNA,gene expression

Henning Urlaub
contact affiliation	Max Planck for Multidisciplinary Sciences Bioanalytical Mass Spectrometry Am Fassberg 11 37077 Gottingen
contact email	henning.urlaub@mpinat.mpg.de
lab head
Alexander Wulf
contact affiliation	Max Planck Institute for Multidisciplinary Sciences
contact email	alexander.wulf@mpinat.mpg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035153
     1. Label: PRIDE project
     2. Name: Npl3 functions in mRNP assembly by recruitment of mRNP components to the transcription site and their transfer onto the mRNA