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Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing.
Open accessAbstract
We have used a site-specific cross-linking strategy to identify RNA and protein factors that interact with the 5' splice site region during mammalian pre-mRNA splicing. Two different pre-mRNA substrates were synthesized with a single 32P-labeled 4-thiouridine residue 2 nucleotides upstream of the 5' splice site. Selective photoactivation of the 4-thiouridine residue after incubation of either substrate under splicing conditions in HeLa nuclear extract resulted in cross-links to the U5 snRNA and the U5 snRNP protein p220. These ATP-dependent interactions occur before the first step of splicing. The U5 snRNA cross-links map to a phylogenetically invariant 9-nucleotide loop sequence and do not require Watson-Crick complementarity to the 5' exon. Cross-links of this position in the pre-mRNA to U1, but not to U2, U4, or U6 snRNAs, were also observed. The kinetics of U1 and U5 cross-link formation are similar, both peaking well before reaction intermediates appear.
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Cited by
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Ørom, U. A., Nielsen, F. C., & Lund, A. H. (2008) · Molecular Cell · doi:10.1016/j.molcel.2008.05.001 · Cited by 1,384
Pre-mRNA splicing and human disease
Faustino, N. A., & Cooper, T. A. (2003) · Genes & Development · doi:10.1101/gad.1048803 · Cited by 1,256
A general two-metal-ion mechanism for catalytic RNA
Steitz, T. A., & Steitz, J. A. (1993) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.90.14.6498 · Cited by 1,171
Self-citationBeyond Secondary Structure: Primary-Sequence Determinants License Pri-miRNA Hairpins for Processing
Auyeung, V. C., Ulitsky, I., McGeary, S. E., & Bartel, D. P. (2013) · Cell · doi:10.1016/j.cell.2013.01.031 · Cited by 434
Dynamic RNA-RNA interactions in the spliceosome
Madhani, H. D., & Guthrie, C. (1994) · Annual Review of Genetics · doi:10.1146/annurev.ge.28.120194.000245 · Cited by 357
Metadata
- Authors
- Wyatt, J. R., Sontheimer, E. J., & Steitz, J. A.
- Venue
- Genes & Development
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 1992-12-01
- Volume
- 6
- Issue
- 12b
- Pages
- 2542-2553
- ISSN
- 0890-9369 (ISSN-L)
- OpenAlex
- W2090643890
- Cited by
- 269
- References
- 0
- Type
- posted content
- Language
- en
- Open access
- diamond
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- CoChat. (2026). Citation check: Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing. https://app.cochat.ai/papers/10.1101/gad.6.12b.2542
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- Wyatt, J. R., Sontheimer, E. J., & Steitz, J. A. (1992). Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing. Genes & Development. https://doi.org/10.1101/gad.6.12b.2542
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