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PreprintCited by 4RNA Research and SplicingRNA and protein synthesis mechanismsGenomics and Chromatin Dynamics

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Real-time tracking of mRNP complex assembly reveals various mechanisms that synergistically enhance translation repression

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Payr, Marco, Meyer, Julia, Geissen, Eva-Maria, Hennig, Janosch, Duss, Olivier·2025·Cell Reports
DOI 10.1101/2025.04.07.647595

Preprint — published as 10.1016/j.celrep.2025.116492

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Abstract

SUMMARY: Protein biosynthesis must be highly regulated to ensure proper spatiotemporal gene expression and thus cellular viability. Translation is often modulated at the initiation stage by RNA binding proteins through either promotion or repression of ribosome recruitment to the mRNA. However, it largely remains unknown how the kinetics of mRNA ribonucleoprotein (mRNP) assembly on untranslated regions (UTRs) relates to its translation regulation activity. Using Sex-lethal (Sxl)-mediated translation repression of msl-2 in female fly dosage compensation as a model system, we show that different mechanisms in mRNP assembly synergistically achieve tight translation repression. Using multi-color single-molecule fluorescence microscopy we show that 1) Sxl targets its binding sites via sliding and double-binding, 2) that Unr recruitment is accelerated over 500-fold by RNA-bound Sxl and 3) that Hrp48 further stabilizes RNA-bound Sxl indirectly via ATP-independent RNA remodeling. Overall, we provide a framework to study how multiple RBPs dynamically cooperate with RNA to achieve function.

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Metadata

Authors
Payr, M., Meyer, J., Geissen, E.-M., Hennig, J., & Duss, O.
Affiliations
  • Heidelberg University (DE)
  • University of Bayreuth (DE)
  • European Molecular Biology Laboratory (DE)
Corresponding author
  • Hennig, Janosch
  • Duss, Olivier
ORCID
Venue
Cell Reports
Publisher
openRxiv
Published
2025-04-10
Volume
44
Issue
11
Pages
116492-116492
ISSN
2211-1247 (ISSN-L)
DOI
10.1101/2025.04.07.647595
OpenAlex
W4415764576
Cited by
4
References
1
Citations by year
2026: 4
FWCI
1.21
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Reports (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
translation repression, RNA-binding proteins, Sex-lethal, Ukrainian People's Republic, dosage compensation, facilitated diffusion, multivalent binding, untranslated regions, single-molecule fluorescence microscopy, Drosophila
MeSH terms
Animals, Drosophila melanogaster, Female, Protein Binding, Ribonucleoproteins, RNA, Messenger, Protein Biosynthesis, RNA-Binding Proteins, Drosophila Proteins

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