Pathogenic Variants in USH1G/SANS Alter Protein Interaction with Pre-RNA Processing Factors PRPF6 and PRPF31 of the Spliceosome
Abstract
:1. Introduction
2. Results
2.1. Binding of SANS to PRPF31 and PRPF6 in the Nucleus Revealed by FRET
2.2. PRPF31 and PRPF6 Interact with Different Regions in SANS CENTn Domain
2.3. In Silico Analysis Predicts Evolutionary Conserved Multi-Conformational Intrinsically Disordered Regions (IRDs) for SANS
2.4. USH-Causing Pathogenic Variants of SANS Show Altered Interaction with PRPFs
3. Discussion
4. Materials and Methods
4.1. DNA Constructs and Primers
4.2. Cloning
4.3. Cell Culture and Cell Lines
4.4. Fluorescence Co-Localization Observation
4.5. Fluorescence Resonance Energy Transfer (FRET) Acceptor Photobleaching Assay
4.6. AlphaFold2-Multimer
4.7. Evolutionary Conservation
4.8. Western Blot
4.9. Additional Bioinformatic Analyses
4.10. Statistics
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Fritze, J.S.; Stiehler, F.F.; Wolfrum, U. Pathogenic Variants in USH1G/SANS Alter Protein Interaction with Pre-RNA Processing Factors PRPF6 and PRPF31 of the Spliceosome. Int. J. Mol. Sci. 2023, 24, 17608. https://doi.org/10.3390/ijms242417608
Fritze JS, Stiehler FF, Wolfrum U. Pathogenic Variants in USH1G/SANS Alter Protein Interaction with Pre-RNA Processing Factors PRPF6 and PRPF31 of the Spliceosome. International Journal of Molecular Sciences. 2023; 24(24):17608. https://doi.org/10.3390/ijms242417608
Chicago/Turabian StyleFritze, Jacques S., Felizitas F. Stiehler, and Uwe Wolfrum. 2023. "Pathogenic Variants in USH1G/SANS Alter Protein Interaction with Pre-RNA Processing Factors PRPF6 and PRPF31 of the Spliceosome" International Journal of Molecular Sciences 24, no. 24: 17608. https://doi.org/10.3390/ijms242417608