Neural Isoforms of Agrin Are Generated by Reduced PTBP1−RNA Interaction Network Spanning the Neuron−Specific Splicing Regions in AGRN
Abstract
:1. Introduction
2. Results
2.1. Alternative Splicing of AGRN Y and Z Exons in Human Cells and Tissues
2.2. Identification of PTBP1 as a Repressor of Alternative Splicing of Y and Z Exons
2.3. Specific Roles of PTBP1−Binding Sites around Y and Z Exons
2.4. The Essential Role of RRM4 in PTBP1 for the Repression of Both Y and Z Exons
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Construction of Minigenes and Expression Vectors for Splicing Analysis
4.3. Transfection of siRNAs and Expression Vectors
4.4. RNA Extraction and RT−PCR
4.5. Immunoblotting
4.6. Tethered Function Assay of PTBP1 and PTBP2
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Bushra, S.; Lin, Y.-N.; Joudaki, A.; Ito, M.; Ohkawara, B.; Ohno, K.; Masuda, A. Neural Isoforms of Agrin Are Generated by Reduced PTBP1−RNA Interaction Network Spanning the Neuron−Specific Splicing Regions in AGRN. Int. J. Mol. Sci. 2023, 24, 7420. https://doi.org/10.3390/ijms24087420
Bushra S, Lin Y-N, Joudaki A, Ito M, Ohkawara B, Ohno K, Masuda A. Neural Isoforms of Agrin Are Generated by Reduced PTBP1−RNA Interaction Network Spanning the Neuron−Specific Splicing Regions in AGRN. International Journal of Molecular Sciences. 2023; 24(8):7420. https://doi.org/10.3390/ijms24087420
Chicago/Turabian StyleBushra, Samira, Ying-Ni Lin, Atefeh Joudaki, Mikako Ito, Bisei Ohkawara, Kinji Ohno, and Akio Masuda. 2023. "Neural Isoforms of Agrin Are Generated by Reduced PTBP1−RNA Interaction Network Spanning the Neuron−Specific Splicing Regions in AGRN" International Journal of Molecular Sciences 24, no. 8: 7420. https://doi.org/10.3390/ijms24087420