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Chicken Y-box proteins chk-YB-1b and chk-YB-2 repress translation by sequence-specific interaction with
S K Swamynathan1, A Nambiar, R V Guntaka
1Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri-Columbia, Columbia, MO 65212, USA.
Abstract:
Y-Box proteins comprise a large family of multifunctional proteins with a wide spectrum of activities in both transcription and translational regulation of gene expression. Earlier, we have reported on the involvement of chk-YB-2 in transcriptional regulation of Rous sarcoma virus long terminal repeats and the involvement of chk-YB-1b in transcriptional regulation of alpha1(I) collagen genes. Here, we have investigated the potential role of chk-YB-2 and chk-YB-1b in RNA metabolism. We report that chk-YB-2 and chk-YB-1b are localized predominantly in the cytoplasm and that they both can bind single-stranded RNA in a sequence-specific and reversible manner. Well-conserved cold-shock domain, N-terminal proline-rich domain and the alternating clusters of acidic and basic amino acids located in the C-terminal ends of these two proteins were all found to be necessary for their RNA-binding ability. Further, we demonstrate that these two proteins inhibit translation in vitro and that binding to RNA is required for this inhibition. The significance of these results is discussed.
Insights
Y-Box proteins chk-YB-2 and chk-YB-1b bind single-stranded RNA and inhibit translation. These findings reveal their crucial role in RNA metabolism and gene expression regulation.
Area of Science:
- Molecular Biology
- Gene Expression Regulation
Background:
- Y-Box proteins are multifunctional, involved in transcription and translation.
- Previous work implicated chk-YB-2 and chk-YB-1b in transcriptional regulation.
Purpose of the Study:
- Investigate the role of chk-YB-2 and chk-YB-1b in RNA metabolism.
- Determine their RNA-binding properties and translational effects.
Main Methods:
- Localization studies to determine cellular compartments.
- RNA-binding assays to assess sequence specificity and reversibility.
- In vitro translation assays to evaluate translational impact.
Main Results:
- chk-YB-2 and chk-YB-1b are primarily cytoplasmic.
- Both proteins bind single-stranded RNA sequence-specifically and reversibly.
- Specific protein domains (cold-shock, proline-rich, C-terminal) are essential for RNA binding.
- RNA binding by these proteins inhibits translation in vitro.
Conclusions:
- chk-YB-2 and chk-YB-1b play significant roles in RNA metabolism.
- Their RNA-binding capacity is critical for translational inhibition.
- These proteins are key regulators of gene expression at the translational level.