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Early commitment of yeast pre-mRNA to the spliceosome pathway.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Legrain, Pierre Séraphin, Bertrand Rosbash, Michael |
| Copyright Year | 1988 |
| Abstract | Pre-mRNA splicing in vitro is preceded by complex formation (spliceosome assembly). U2 small nuclear RNA (snRNA) is found in the earliest form of the spliceosome detected by native gel electrophoresis, both in Saccharomyces cerevisiae and in metazoan extracts. To examine the requirements for the formation of this early complex (band III) in yeast extracts, we cleaved the U2 snRNA by oligonucleotide-directed RNase H digestion. U2 snRNA depletion by this means inhibits both splicing and band III formation. Using this depleted extract, we were able to design a chase experiment which shows that a pre-mRNA substrate is committed to the spliceosome assembly pathway in the absence of functional U2 snRNP. Interactions occurring during the commitment step are highly resistant to the addition of an excess of unlabeled substrate and require little or no ATP. Sequence requirements for this commitment step have been analyzed by competition experiments with deletion mutants: both the 5' splice site consensus sequence and the branch point TACTAAC box sequence are necessary. These experiments strongly suggest that the initial assembly process requires a trans-acting factor(s) (RNA and/or proteins) that recognizes and stably binds to the two consensus sequences of the pre-mRNA prior to U2 snRNP binding. |
| Starting Page | 384 |
| Ending Page | 387 |
| Page Count | 4 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://www.bio.brandeis.edu/rosbash-lab/PublishedPapers/1992-1996/1987-1991/LeGrain-MCB-1988.pdf |
| Alternate Webpage(s) | http://www.bio.brandeis.edu/rosbashlab/PublishedPapers/1992-1996/1987-1991/LeGrain-MCB-1988.pdf |
| PubMed reference number | 3065622v1 |
| Volume Number | 8 |
| Issue Number | 9 |
| Journal | Molecular and cellular biology |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Cleaved cell Deletion Mutation Gel Electrophoresis (lab technique) Pierre Robin Syndrome RNA Splicing Ribonucleoprotein, U2 Small Nuclear Small Nuclear RNA Spliceosome Assembly Pathway Spliceosomes interferon alfacon-1 mRNA Precursor mutant negative regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay |
| Content Type | Text |
| Resource Type | Article |
