Genome-wide discovery of human splicing branchpoints

Mercer, Tim R., Clark, Michael B., Andersen, Stacey B., Brunck, Marion E., Haerty, Wilfried ORCID:, Crawford, Joanna, Taft, Ryan J., Nielsen, Lars K., Dinger, Marcel E. and Mattick, John S. (2015) Genome-wide discovery of human splicing branchpoints. Genome Research, 25 (2). pp. 290-303. ISSN 1088-9051

[thumbnail of GenomeRes-2015-Mercer-290-303]
PDF (GenomeRes-2015-Mercer-290-303) - Published Version
Available under License Creative Commons Attribution Non-commercial.

Download (1MB) | Preview


During the splicing reaction, the 59 intron end is joined to the branchpoint nucleotide, selecting the next exon to incorporate into the mature RNA and forming an intron lariat, which is excised. Despite a critical role in gene splicing, the locations and features of human splicing branchpoints are largely unknown. We use exoribonuclease digestion and targeted RNA-sequencing to enrich for sequences that traverse the lariat junction, by split and inverted alignment, reveal the branchpoint. We identify 59,359 high-confidence human branchpoints in >10,000 genes, providing a first map of splicing branchpoints in the human genome. Branchpoints are predominantly adenosine, highly conserved, and closely distributed to the 3 ′ splice site. Analysis of human branchpoints reveals numerous novel features, including distinct features of branchpoints for alternatively spliced exons and a family of conserved sequence motifs overlapping branchpoints we term B-boxes, which exhibit maximal nucleotide diversity while maintaining interactions with the keto-rich U2 snRNA. Different B-box motifs exhibit divergent usage in vertebrate lineages and associate with other splicing elements and distinct intron-exon architectures, suggesting integration within a broader regulatory splicing code. Lastly, although branchpoints are refractory to common mutational processes and genetic variation, mutations occurring at branchpoint nucleotides are enriched for disease associations.

Item Type: Article
Uncontrolled Keywords: genetics,genetics(clinical),sdg 3 - good health and well-being ,/dk/atira/pure/subjectarea/asjc/1300/1311
Faculty \ School: Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Medicine and Health Sciences > Research Centres > Norwich Institute for Healthy Aging
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 19 Feb 2021 01:04
Last Modified: 21 Apr 2023 00:58
DOI: 10.1101/gr.182899.114

Actions (login required)

View Item View Item