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PMID: 10691733 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The carboxyl terminus of vertebrate poly(A) polymerase interacts with U2AF 65 to couple 3'-end processing and splicing.

Genes & development ·Vol. 14 ·No. 4 ·2000-02-15 ·Pages 403-13

Vagner S, Vagner C, Mattaj IW

Abstract

Although it has been established that the processing factors involved in pre-mRNA splicing and 3'-end formation can influence each other positively, the molecular basis of this coupling interaction was not known. Stimulation of pre-mRNA splicing by an adjacent cis-linked cleavage and polyadenylation site in HeLa cell nuclear extract is shown to occur at an early step in splicing, the binding of U2AF 65 to the pyrimidine tract of the intron 3' splice site. The carboxyl terminus of poly(A) polymerase (PAP) previously has been implicated indirectly in the coupling process. We demonstrate that a fusion protein containing the 20 carboxy-terminal amino acids of PAP, when tethered downstream of an intron, increases splicing efficiency and, like the entire 3'-end formation machinery, stimulates U2AF 65 binding to the intron. The carboxy-terminal domain of PAP makes a direct and specific interaction with residues 17-47 of U2AF 65, implicating this interaction in the coupling of splicing and 3'-end formation.

MeSH Terms
3' Untranslated Regions/metabolism Amino Acid Sequence HeLa Cells/metabolism Humans Introns Macromolecular Substances Molecular Sequence Data Neoplasm Proteins/metabolism Nuclear Proteins Polynucleotide Adenylyltransferase/metabolism Protein Binding RNA Precursors/metabolism RNA Processing, Post-Transcriptional/physiology RNA Splicing/physiology RNA, Neoplasm/metabolism Regulatory Sequences, Nucleic Acid Ribonucleoproteins/metabolism Ribonucleoproteins, Small Nuclear/metabolism Splicing Factor U2AF
Chemicals
3' Untranslated Regions Macromolecular Substances Neoplasm Proteins Nuclear Proteins RNA Precursors RNA, Neoplasm Ribonucleoproteins Ribonucleoproteins, Small Nuclear Splicing Factor U2AF U2AF2 protein, human Polynucleotide Adenylyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Vagner S
European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.
Vagner C
Mattaj I W
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
Published
2000-02-15
Pages
403-13
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316384
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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