ArticleRNA (New York, N.Y.)2000
Characterization of human RNA splice signals by iterative functional selection of splice sites.
Article in RNA (New York, N.Y.), 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Dichotomous splicing signals in exon flanks.Genome research · 2005Article
- A novel intronic cis element, ISE/ISS-3, regulates rat fibroblast growth factor receptor 2 splicing through activation of an upstream exon and repression of a downstream exon containing a noncanonical branch point sequence.Molecular and cellular biology · 2005Article
- Sequence information for the splicing of human pre-mRNA identified by support vector machine classification.Genome research · 2003Article
- Genome-wide prediction, display and refinement of binding sites with information theory-based models.BMC bioinformatics · 2003Article
- Murine serine proteases MASP-1 and MASP-3, components of the lectin pathway activation complex of complement, are encoded by a single structural gene.Genes and immunity · 2003Article
- Sodium channels SCN1A, SCN2A and SCN3A in familial autism.Molecular psychiatry · 2003Article
- Defining a 5' splice site by functional selection in the presence and absence of U1 snRNA 5' end.RNA (New York, N.Y.) · 2002Article
- Mutational analysis of a plant branchpoint and polypyrimidine tract required for constitutive splicing of a mini-exon.RNA (New York, N.Y.) · 2002Article
- Control of branch-site choice by a group II intron.The EMBO journal · 2001Article
- Functional selection of splicing enhancers that stimulate trans-splicing in vitro.RNA (New York, N.Y.) · 2001Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
An iterative in vitro splicing strategy was employed to select for optimal 3' splicing signals from a pool of pre-mRNAs containing randomized regions. Selection of functional branchpoint sequences in HeLa cell nuclear extract yielded a sequence motif that evolved from UAA after one round of splicing toward a UACUAAC consensus after seven rounds. A significant part of the selected sequences contained a conserved AAUAAAG motif that proved to be functional both as a polyadenylation signal and a branch site in a competitive manner. Characterization of the branchpoint in these clones to either the upstream or downstream adenosines of the AAUAAAG sequence revealed that the branching process proceeded efficiently but quite promiscuously. Surprisingly, the conserved guanosine, adjacent to the common AAUAAA polyadenylation motif, was found to be required only for polyadenylation. In an independent experiment, sequences surrounding an optimal branchpoint sequence were selected from two randomized 20-nt regions. The clones selected after six rounds of splicing revealed an extended polypyrimidine tract with a high frequency of UCCU motifs and a highly conserved YAG sequence in the extreme 3' end of the randomized insert. Mutating the 3' terminal guanosine of the intron strongly affects complex A formation, implying that the invariant AG is recognized early in spliceosome assembly.
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