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Παρασκευή 12 Μαΐου 2017

Exploring the Impact of Cleavage and Polyadenylation Factors on Pre-mRNA Splicing Across Eukaryotes

In human, mouse and Drosophila, the spliceosomal complex U1 snRNP (U1) protects transcripts from premature cleavage and polyadenylation at proximal intronic polyadenylation signals (PAS). These U1-mediated effects preserve transcription integrity and are known as telescripting. The watchtower role of U1 throughout transcription is clear. What is less clear is whether cleavage and polyadenylation factors (CPFs) are simply patrolled or if they may actively antagonize U1 recruitment. In addressing this question, we found that in the introns of human, mouse and Drosophila and of 14 other eukaryotes, including multi- and single-celled species, the conserved AATAAA PAS — a major target for CPFs — is selected against. This selective pressure, approximated using DNA strand asymmetry, is detected for peripheral and internal introns alike. It is surprisingly more pronounced within — rather than outside — the action range of telescripting and particularly intense in the vicinity of weak 5' splice sites. Our study uncovers a novel feature of eukaryotic genes: that the AATAAA PAS is universally counter-selected in spliceosomal introns. This pattern implies that CPFs may attempt to access introns at any time during transcription. However, natural selection operates to minimize this access. By corroborating and extending previous work, our study further indicates that CPF access to intronic PASs might perturb the recruitment of U1 to the adjacent 5' splice sites. These results open the possibility that CPFs may impact the splicing process across eukaryotes.



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